Coomassie Blue Research Articles

SNP rs4971059 predisposes to breast carcinogenesis and chemoresistance via TRIM46-mediated HDAC1 degradation.

Identification of the driving force behind malignant transformation holds the promise to combat the relapse and therapeutic resistance of cancer. We report here that the single nucleotide polymorphism (SNP) rs4971059, one of 65 new breast cancer risk loci identified in a recent genome-wide association study (GWAS), functions as an active enhancer of TRIM46 expression. Recreating the G-to-A polymorphic switch caused by the SNP via CRISPR/Cas9-mediated homologous recombination leads to an overt upregulation of TRIM46. We find that TRIM46 is a ubiquitin ligase that targets histone deacetylase HDAC1 for ubiquitination and degradation and that the TRIM46-HDAC1 axis regulates a panel of genes, including ones critically involved in DNA replication and repair. Consequently, TRIM46 promotes breast cancer cell proliferation and chemoresistance invitro and accelerates tumor growth invivo. Moreover, TRIM46 is frequently overexpressed in breast carcinomas, and its expression is correlated with lower HDAC1 expression, higher histological grades, and worse prognosis of the patients. Together, our study links SNP rs4971059 to replication and to breast carcinogenesis and chemoresistance and support thepursuit of TRIM46 as a potential target for breast cancer intervention.

Green synthesis of silver nanoparticles by Exiguobacterium profundum isolated from carpet industry effluent

Green synthesis of silver nanoparticles using microorganisms has received profound interest because of their potential to synthesize nanoparticles of various size, shape and morphology. The main objective of the current study was to evaluate the synthesis of silver nanoparticles by a bacterial strain, Exiguobacterium profundum with potential dye degrading ability isolated from carpet industry effluent. The strain was tested for its ability to degrade various dyes including Methylene Blue, Malachite Green, Congo Red, Crystal Violet, Coomassie Brilliant Blue, Methyl Red, Sudan Black. Treatment of the bacteria with 1 mM AgNO3, showed its ability to form silver nanoparticles. This was confirmed by visual observation and UV–Vis absorption at 450 nm. Further characterization of nanoparticles by Scanning electron microscopy confirmed the size of silver nanoparticles in 20-40 nm range. Therefore, the present study is a demonstration of an efficient synthesis of stable silver nanoparticles using Exiguobacterium profundum which could be utilized in dye degradation thereby reducing the pollution from industrial effluents.

Zinc oxide quantum dots: Confinement size, photophysical and tunning optical properties effect on photodecontamination of industrial organic pollutants

Confinement size effect and tunning optical properties of Zinc oxide quantum dots (ZDs) observed via modified wet chemical preparation method. The purity, high crystallinity, and elongated structure of ZDs investigated by XRD and TEM tests. The crystallized size values, the bandgap energy, and the active surface area for ZD1 and ZD2 determined based on the TEMs, DRSs, and EBTs are 2.9 and 3.6 nm, 3.57 eV and 3.54 eV, and 312.10 m2/g and 278.05 m2/g, respectively. Absorption and fluorescence spectra shifted for ZDs from 340 to 315 nm and from 530 to 500 nm, respectively, due to its confinement effect of their small sizes. The prepared samples revealed that the photocatalytic activity of ZD1 is higher than ZD2 by 20%. Photocatalytic activity of ZDs by the photodegradation process of Coomassie Brilliant Blue R dye as an industrial organic pollutant evaluated (used in textile and dyeing manufactories) by xenon photoreactor and solar photocatalytic processes. The observed results have shown that photocatalytic efficiency increases with crystallite size decreasing. And, the variation between the photocatalytic activity of ZDs prepared samples by the solar photocatalytic process is higher than that observed by the Xenon photoreactor. And their photodegradation rates values are lower than that obtained by the Xenon photoreactor. A similar finding for the recycling process after nine reptation times of using ZDs during all photodegradation processes signified obtained.

Photocatalytic Synthesis of Benzimidazoles on Heterogenized Coomassie Brilliant Blue

Key words photocatalysis - tungsten nanoparticles - heterogeneous catalysis - benzimidazoles

Novel Poly(ionic liquid) Augmented Membranes for Unconventional Aqueous Phase Applications in Fractionation of Dyes and Sugar.

Poly(ionic) liquid (PIL) augmented membranes were fabricated through self-polymerization of 2-vinyl pyridine and 4-vinyl pyridine followed by dopamine triggered polymerization and bridging with inert polyamide support. The resulting membranes acquired a positive surface charge with a high degree of hydrophilicity. Fourier transformed Infra-red (FTIR) and Energy dispersive X-ray (EDX) spectroscopic investigation revealed the successful augmentation of PIL surface layer, whereas surface morphology was investigated through scanning electron microscopy (SEM) imaging. This manuscript demonstrates pi electron-induced separation of dyes with the trend in permeability: Coomassie Brilliant Blue G (CBBHG) > Remazol Brilliant Blue R (RBBR) > Eichrome Black T (EBT) > Congo Red (CR). CBBG exhibited extended conjugation over large aromatic domain. RBBR and EBT were associated withtheelectron-donating -NH2 group and electron-withdrawing -NO2 group, respectively, hence pi electron density on aromatic ring varied. The steric repulsion between two pairs of ortho hydrogens (Hs) in biphenyl moieties of CR resulted in deviation of planarity and hence aromaticity leading to the lowest permeability. The sugar fractionation followed the trend: Galactose > Mannose > Fructose > Glucose > Xylose. More hydroxyl (-OH) groups in sugars and their conformational alignment in the same direction, exhibited more lone pair of electrons leading to more interaction with PIL and hence better permeability. Pentose showed poorer permeation than hexose, whereas aldose showed better permeation than ketose.

Dye-Loaded Polymersome-Based Lateral Flow Assay: Rational Design of a COVID-19 Testing Platform by Repurposing SARS-CoV-2 Antibody Cocktail and Antigens Obtained from Positive Human Samples.

The global pandemic of COVID-19 continues to be an important threat, especially with the fast transmission rate observed after the discovery of novel mutations. In this perspective, prompt diagnosis requires massive economical and human resources to mitigate the disease. The current study proposes a rational design of a colorimetric lateral flow immunoassay (LFA) based on the repurposing of human samples to produce COVID-19-specific antigens and antibodies in combination with a novel dye-loaded polymersome for naked-eye detection. A group of 121 human samples (61 serums and 60 nasal swabs) were obtained and analyzed by RT-PCR and ELISA. Pooled samples were used to purify antibodies using affinity chromatography, while antigens were purified via magnetic nanoparticles-based affinity. The purified proteins were confirmed for their specificity to COVID-19 via commercial LFA, ELISA, and electrochemical tests in addition to sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Polymersomes were prepared using methoxy polyethylene glycol-b-polycaprolactone (mPEG-b-PCL) diblock copolymers and loaded with a Coomassie Blue dye. The polymersomes were then functionalized with the purified antibodies and applied for the preparation of two types of LFA (antigen test and antibody test). Overall, the proposed diagnostic tests demonstrated 93 and 92.2% sensitivity for antigen and antibody tests, respectively. The repeatability (92–94%) and reproducibility (96–98%) of the tests highlight the potential of the proposed LFA. The LFA test was also analyzed for stability, and after 4 weeks, 91–97% correct diagnosis was observed. The current LFA platform is a valuable assay that has great economical and analytical potential for widespread applications.

A critical green biosynthesis of novel CuO/C porous nanocomposite via the aqueous leaf extract of Ficus religiosa and their antimicrobial, antioxidant, and adsorption properties

Herein, we report the biosynthesis of Copper oxide/Carbon (CuO/C) nanocomposite via the aqueous decoction of Ficus religiosa leaves as a capping factor, an origin of carbon and a reducing agent. CuSO4.5H2O solution is adopted as the precursor for synthesizing CuO/C nanocomposite. The biosynthesized nanocomposite was characterized by using UV-Visible, Micro Raman, XPS, FTIR and FESEM. All these characterization results established that the fabricated substance is a nanocomposite of carbon and CuO. The porous structure of nanocomposite was accompanied by effective adsorption of organic dyes Coomassie Brilliant Blue (CBB), Methylene Blue (MB) and Congo Red (CR) from water. The effect of time, temperature, the concentration of dyes and mass of nanocomposite were investigated. The data of adsorption for all three dyes agreed properly with Pseudo second-order kinetics and Langmuir isotherm models. The adsorption capacities of the nanocomposite for CR, MB and CBB dyes are evaluated to be 20 mg/g, 7.582 mg/g and 15.23 mg/g respectively In addition, the nanocomposite exhibited strong antimicrobial performance and antioxidant activity. Conclusively, it can be emphasized that the CuO/C nanocomposites could be useful for various industrial and biomedical applications.

Multifunctional ZnO nanoparticles bio-fabricated from Canna indica L. flowers for seed germination, adsorption, and photocatalytic degradation of organic dyes

The potential of green nanomaterials for environmental and agricultural fields is emerging due to their biocompatible, eco-friendly, and cost-effective performance. We report the use of Canna indica flowers extract as new capping and stabilizing source to bio-fabricate ZnO nanoparticles (ZnO NPs for dyes removal, seed germination. ZnO NPs was biosynthesized by ultrasound-assisted alkaline-free route to reach the critical green strategy. The physicochemical findings of ZnO revealed small crystallite size (27.82 nm), sufficient band-gap energy (3.08 eV), and diverse functional groups. Minimum‑run resolution IV approach found the most pivotal factors influencing on removal of Coomassie Brilliant Blue G-250. Uptake studies pointed out that pseudo second-order, and Langmuir were the best fitted models. Dye molecules behaved monolayer adsorption on ZnO surface layers, and controlled by chemisorption. Natural solar light was used as effective source for photocatalytic degradation of methylene blue (94.23% of removal and 31.09 mg/g of uptake capacity). Compared with H2O and ZnSO4, ZnO NPs positively affected the growth of shoot and root lengths (10.2–27.8%) of bean seedlings in most cases. ZnO acts an agrochemical for boosting weight gain, and germination ratio. This study may be promising for developing the recyclable, multifunctional ZnO nanoparticles for environmental and agricultural applications.

Facile fabrication of polymeric quaternary ammonium salt hydrogel by radiation for dyes removal from aqueous solution

The effluents containing various dyes from the process industries raise esthetic problems and cause toxic impacts on aquatic and human life. Thus, the dye effluent must be adequately treated before being discharged into water-bodies. A cellulose-derived quaternary ammonium salt was treated by radiation to form hydrogel structure. The radiation dose effects on gel fraction and swelling properties were investigated. The hydrogel structure was characterized by IR and SEM analyses. The Zeta potential values at different pH were all greater than +30 mV, which indicating that the hydrogel possessed enough electrostatic attraction force to anionic ions. The adsorption behaviors of the polymeric quaternary ammonium salts hydrogel (PQH) towards methyl orange (MO), Congo red (CR), and Coomassie brilliant blue (CBB) are discussed. The pseudo-second-order mode well fitted the adsorption kinetics for all three dyes. The increasing order of the initial adsorption rate h0 was MO > CR > CBB. The Langmuir model well described the adsorption isotherms were well described by with the maximum adsorption capacity for MO, CR, and CBB being 628.93, 427.35, and 617.28 mg/g, respectively. HCl was used to regenerate the dye-loaded-hydrogel. The adsorption-desorption test demonstrated the good reusability of this hydrogel. Also, the PQH has selective adsorption of MO from RhB/MO mixing solution.

Optimization of protein extraction and ELISA immunodetection from protein-based paint models with mesoporous silica nanoparticles and MCM41

Protein-based biological materials such as albumin, casein and collagen are found in various cultural heritage (CH) artefacts. This study focuses on the study of protein binders from easel paintings media. Proteins have complex structures which are difficult to identify with non-invasive spectroscopic methods (FT-IR, Raman, UV). Immunoassays such as ELISA determine the protein’s source of origin which is necessary for art objects. To increase the detection and identification of proteins by immunoassays, the efficiency of micro-extraction of proteins from heritage materials is a crucial step. Extractions mediated by cycles of orbital agitation and ultrasonic radiation give the possibility to extract proteins from easel painting sample. In this work, protein-based paint models coupled with silica nanoparticles were used for micro-extraction. Nanoparticles possess high surface-to-volume ratios that can attach bioactive molecules such as proteins and increase the total protein recovered from microsamples. Protein extracts were quantified with Bradford Assay in the presence of Coomassie blue. The protein recovery results were statistically computed, and the SPSS analysis shows significant (p < 0.05) increase in protein recovery, above 1.3 times for NPSiO2 and above 1.6 times for MCM-41. The statistical data shows evidence that silica nanoparticles intensify the total protein recovered from paint microsamples. Finally, ELISA was realized on the protein extracts to verify and compare the immunodetection of protein from the paint models with and without the use of silica nanoparticles.

Adsorption of Malachite Green by extracellular polymeric substance of Lysinibacillus sp. SS1: kinetics and isotherms

Use of novel biological materials as adsorbents for removal of xenobiotics is gaining significance owing to their exceptional advantages. An extracellular polymeric substance (EPS) produced by Lysinibacillus sp. SS1 had rough porous surface as observed by SEM analysis. Adsorption ability of EPS was estimated against various textile dyes such as Malachite Green (MG), Methyl Orange, Congo Red and Coomassie Blue. About 82% of MG (100 mg/L) was adsorbed onto 2.5 mg EPS within 30 min. Effect of MG concentration, EPS weight, agitation speed and incubation time on adsorption, studied by one factor at a time approach, revealed that adsorption was influenced by all factors. Maximum adsorption of 99.01 ± 0.61% was achieved at 100 mg/L MG, 10 mg EPS, 120 RPM in 75 min with maximum adsorption capacity of 247.5 mg/g. Kinetics was affected by MG and EPS amounts, with shift from pseudo first to pseudo second order with increase in concentration. Adsorption of MG by EPS of Lysinibacillus sp. SS1 was identified as unilayer chemisorption as it followed Langmuir isotherm with maximum adsorption capacity (Qm) of 178.57 mg/g (R2 = 0.9889). This is the first report on potential of EPS produced by Lysinibacillus sp. SS1 as novel biodegradable adsorbent with high efficacy of MG removal from aqueous solutions.

Centella asiatica crop residue fabricated silver nanoparticles as potent antioxidant agents in photo-catalytic degradation of hazardous dyes

Centella asiatica (CA) is a valuable crop for herbal, cosmetic, and medicinal industries due to its rich triterpenoids, and saponins in leaves. The CA petioles, stem, and runners (CAP) are considered as crop residue and in this study for the first time CAP extract was used for Photo-mediated synthesis of silver nanoparticles (SNPs). Further, the synthesis process was optimized using response surface methodology. Synthesized SNPs were highly stable (-30.8 eV), spherical, and cube shape with a crystallite size of 22.83 nm. Phytochemical screening, FTIR, and Raman studies confirmed the presence of bioactive functional groups on CAP SNPs. The CAP extract and its SNPs confirmed higher antioxidant activity compared to standard (Ascorbic acid). The results of hazardous dye degradation studies showed that percentage dye degradation was higher in methyl orange (1 min; 97.54%) followed by Congo red (4 min; 90.86%), methyl red (10 min; 97.66%), and Coomassie Brilliant Blue R-250 (30 min; 93.28%) with rate constant of 0.062/sec, 0.00062/sec, 0.0099/sec, and 0.0015/sec respectively.

The comfort properties of cosmeto-textiles functionalized with protein-based nanoemulsions encapsulating Vitamin-E

ABSTRACT The comfort properties of a textile cannot be dissociated from its function. Considering this premise, we evaluated the physical and mechanical performance of a previously developed cosmeto-textile composed of cotton/elastane, functionalized with bovine serum albumin (BSA) and BSA/silk fibroin (BSA/SF) nanoemulsions, encapsulating α-tocopherol (vitamin E). The surface functionalization of cotton fabrics with the nanoemulsions was confirmed by fabric staining with Coomassie Brilliant Blue R250 and by FTIR-ATR. The comfort properties including wetting, stiffness, stretching, draping ability, electrical resistance, and air and vapor permeability were evaluated. The results revealed that the functionalization of cotton fabrics with the developed protein-based nanoemulsions did not disturb the comfort properties of the textile material nor its functionality. This gives a good indication of the potential of the developed breathable cosmeto-textile to be used in contact with the human body with acceptable performance.

Next-Generation Dried Blood Spot Samplers for Protein Analysis: Describing Trypsin-Modified Smart Sampling Paper

This paper describes smart sampling paper to be used for bottom-up protein analysis. Four different manners to immobilize trypsin on cellulose were evaluated. Untreated paper, potassium-periodate-functionalized paper (with and without post-immobilization reduction) and 2-hydroxyethyl methacrylate (HEMA)/2-vinyl-4,4-dimethylazlactone (VDM)-functionalized paper were all used to immobilize trypsin. For the evaluation, Coomassie Brilliant Blue staining of proteins on paper and the BAEE trypsin activity assay needed to be modified. These methods allowed, together with data from mass spectrometric analysis of cytochrome C digestions, us to acquire fundamental insight into protein binding, and trypsin action and activity on paper. All functionalized discs bind more protein than the untreated discs. Protein binding to functionalized discs is based on both adsorption and covalent binding. Trypsin immobilized on potassium-periodate-functionalized discs exhibits the highest trypsin activity when using cytochrome C as substrate. It is proven that it is trypsin attached to paper (and not desorbed trypsin) which is responsible for the enzyme activity. The use of discs on complex biological samples shows that all functionalized discs are able to digest diluted serum; for the best-performing disc, HEMA-VDM functionalized, up to 200 high-confidence proteins are qualified, showing its potential.

Preparation of raw oyster shell for removal of coomassie brilliant blue R-250 dye from aqueous solution

The removal of dye from textile wastewater effluent has become an important process nowadays. In this study, raw oyster shell was used as adsorbent for removal of Coomassie Brilliant Blue R-250 (CBB) dye by using the adsorption method in order to reduce the amount of waste oyster shells in waters and landfills which can cause pollution. Various parameters such as adsorbent size, adsorbent dosage, initial dye concentration, contact time and pH were investigated to study the potential of oyster shell as an adsorbent. The optimum adsorption was obtained at adsorbent size of 75 µm and 2.3 g of adsorbent dosage at 50 mg/L concentration of CBB dye. Optimum contact time of the dye adsorption was determined at 9 hours and the most effective condition for CBB dye removal was at pH 2. The removal efficiency was achieved at 99.64%. Thus, this study shows potential as an economical and environment friendly dye removal process for wastewater treatment.

Fibrin – Fibrinogen scaffold structural modification by phytochemicals and phytoproteases contained in an aqueous extract of Ageratum conyzoides Linn.

Based on mechanisms of fibrin clot polymerization and dissolution, it is possible to modulate fibrin formation and removal. Ageratum conyzoides Linn. (Asteraceae) is an annual herb with a long history of traditional medicine. There is high variability in the secondary metabolites of this plant which include flavonoids, and these molecules belong to a class of serine proteases inhibitors. Several plant enzymes belonging to the classes of serine proteases were observed to be active on the cascade of coagulation pathways. The aim of this study was to observe if even Ageratum conyzoides Linn. aqueous leaves extract contained proteases which could structurally modify the fibrin clot formation. To prepare plant extracts, dry leaves of the plant were extracted with distilled water. Fibrin gels were prepared by mixtures containing fibrinogen and thrombin with or without extract. Fibrin networks were disrupted by a denaturation buffer. Samples were deposited in 8% polyacrylamide gel and Coomassie blue was used to reveal migration. Our extract contained phytochemicals class flavonoids which are thrombin inhibitors. But our results support the evidence that the same extract contained plant serine proteases, specifically a fibrinogenase which hydrolyzed fibrinogen but not like thrombin.Keywords: Fibrin/Fibrinogen, structural modification, Ageratum conyzoides Linn., phytoproteases.

A new family of Ln-BTC-AC-FM framework intelligent materials: Precise synthesis, structure and characterization for fluorescence detecting of UO22+ and adsorbing dyes

The unique fluorescence properties of rare earth ions and subtle interactions with organic ligands make the target lanthanide metal-organic frameworks show potential development prospects in many fields. Based on this situation, a series of new lanthanide metal-organic frameworks Ln-BTC-AC-FM: (CH3)2NH2[Ln2(BTC)(AC)3(FM)], (Ln=Pr (1), Ce (2), Nd (3), Eu (4), Sm (5), Gd (6), Tb (7), Ho (8), Er (9) and Yb (10), H3BTC=1,3,5-benzenetricarboxylic acid) were synthesized through the reaction of triangular polycarboxylic acid ligand (H3BTC) and rare earth nitrate Ln(NO3)3·6H2O with mixed formic acid (FM) and glacial acetic acid (AC) system. Due to this new type of Ln-BTC-AC-FM MOFs have unique and excellent fluorescence properties, we selected (CH3)2NH2[Eu2(BTC)(AC)3(FM)] (4) to explore its detection performance for UO22+ in aqueous solution. The study found that 4 is a high-efficiency fluorescent probe, which has a good quenching effect on uranium solution, and the quenching rate can reach 98.01%. At the same time, 4 has a good fluorescent response to UO22+, with a higher KSV value of 8.56 × 103 M−1 and a lower detection limit of 4.12 μM. In addition, we also studied the adsorption properties of this series of Ln-BTC-AC-FM MOFs for dyes because of their novel three-dimensional microporous structures. The results showed that 4 has a good adsorption effect on Coomassie Brilliant Blue (CBB) and Eosin. Moreover, it was found that the dye adsorption process was proved to be a suitable pseudo-second-order kinetic model.

Ultrasound-irradiated synthesis of 3-mercaptopropyl trimethoxysilane-modified hydroxyapatite derived from fish-scale residues followed by ultrasound-assisted organic dyes removal

We report a novel method for the synthesis of 3-mercaptopropyl trimethoxysilane-modified hydroxyapatite (FHAP-SH) derived from fish-scale residues by using ultrasound irradiation. Scanning electron microscopy, transmission electron microscopy, energy-dispersive spectroscopy, X-ray diffraction, and Fourier transform infrared spectroscopy were used for the FHAP-SH characterization. Then, the organic dye adsorption on the FHAP-SH was monitored through an ultrasound process. After the dye removal optimization, significant improvements were observed in the maximum adsorption capacities for Congo Red (CR, 500 mg g−1), Coomassie Brilliant Blue G 250 (CB, 235 mg g−1), and Malachite Green (MG, 625 mg g−1). The adsorption behaviors of these dyes were fitted by using the Langmuir isotherm model with a high coefficient of determination values ranging from 0.9985 to 0.9969. The adsorption of the three dyes onto FHAP-SH was an endothermic process based on the adsorption thermodynamics model, while the adsorption kinetics analysis of the dyes presented a good alignment with the pseudo-second-order kinetics. The FHAP-SH exhibits a remarkably high adsorption capacity, is inexpensive, and fulfills the ecofriendly requirements of dye wastewater treatment, especially in the textile industry.

Lateral flow immunoassay for furazolidone point-of-care testing: Cater to the call of saving time, labor, and cost by coomassie brilliant blue labeling

Furazolidone (FZD) and its metabolite called 3-amino-2-oxazolidinone (AOZ) would induce carcinogenic and mutagenic effects to human. In this work, to develop a novel, stable, and simple point of care testing (POCT) with a potential to social applied for FZD detection, we utilized the aspect of protein staining of coomassie brilliant blue (CBB) to exploit a new CBB-LFIA strategy free of NPs. Only one mixing step is needed during the probe manufacturing process, which requires just 2 h and is a great time saving strategy compared with other methods (requiring 4–33 h for probe preparation). Besides, the cost of CBB-LFIA is 300 times lesser than other LFIA with respect to obtaining the label. The developed CBB-LFIA was successfully applied to detect AOZ with a detection limit of 2 ng mL−1, without any influence from other potential interfering compounds. The proposed CBB-LFIA exhibited prominent practical application, and possesses considerable utilization potential in the related field.

Alterations ofhousekeepingproteins in human aged and diseased hearts.

Pathological remodeling includes alterations of ion channel function and calcium homeostasis and ultimately cardiac maladaptive function during the process of disease development. Biochemical assays are important approaches for assessing protein abundance and post-translational modificationof ion channels. Several housekeeping proteins are commonly used as internal controls to minimize loading variabilitiesin immunoblotting protein assays. Yet, emerging evidence suggests that some housekeeping proteins may be abnormally altered under certain pathological conditions. However, alterations of housekeeping proteins in aged and diseased human hearts remain unclear. In the current study, immunoblotting was applied to measure three commonly used housekeeping proteins (β-actin, calsequestrin, and GAPDH) in well-procured human right atria (RA) and left ventricles (LV) from diabetic, heart failure, and aged human organ donors. Linear regression analysis suggested that the amounts of linearly loaded total proteins and quantified intensity of total proteins from either Ponceau S (PS) blot-stained or Coomassie Blue (CB) gel-stained images were highly correlated. Thus, all immunoblotting data were normalized with quantitative CB or PS data to calibrate potential loading variabilities. In the human heart, β-actin was reduced in diabetic RA and LV, while GAPDH was altered in aged and diabetic RA but not LV. Calsequestrin, an important Ca2+ regulatory protein, was significantly changed in aged, diabetic, and ischemic failing hearts. Intriguingly, expression levels of all three proteins were unchanged in non-ischemic failing human LV. Overall, alterations of human housekeeping proteins are heart chamber specific and disease context dependent. The choice of immunoblotting loading controls should be carefully evaluated. Usage of CB or PS total protein analysis could be a viable alternative approach for some complicated pathological specimens.