BCA Protein Quantitative Detection Kit

Descriptions

BCA Kit는 5% SDS, 5% Triton x-100, 5% Tween-20, 60, 80 등과 같은 detergent에 영향을 받지 않기에 이 것들의 농도가 높은 샘플을 정량하는데 매우 유용합니다. 그러나 고농도 환원제와 chelating agent에는 영향을 받습니다. 따라서 샘플에 EGTA가 없는지, EDT의 농도가 10mM 미만인지, DTT나 beta-mercaptoethanol의 농도가 1mM미만인지 확인해야합니다.

Storage and Handling Conditions

The whole reagent can be transported at room temperature; Protein standard (BSA) stored at 4℃, valid for 12 months. The protein standard solution should be stored at -20℃ and used within 6 months. The remaining reagents are stored at room temperature, valid for 12 months.

Product Information

Component

Assay Protocol / Procedures

1. Preparation of protein standard storage solution: 1 mL protein standard diluent solution was added to protein standard tube (BSA), and 25 mg protein standard was completely dissolved to obtain protein standard storage solution with a concentration of 25 mg/mL. The standard protein storage solution can be preserved for a long time at -20℃； 

2. Preparation of protein standard working solution: 25 mg/ml protein standard storage solution is diluted 50 times with PBS or normal saline to obtain protein standard working solution with a final concentration of 0.5 mg/ml. Pay attention to the 10 times gradient method for dilution to ensure accuracy. 

3. Standard curve (Microplate Procedure): use the following table as a guide to prepare a set of protein standards:

1. Preparation of test-sample: the test-sample is diluted appropriately (through the pre-test detection, the protein concentration of the test-sample is within the range of the standard curve to ensure that the detection results are reliable), and pipette 20 µL of each test-sample into the 96-well plates. The test-sample and the protein standard working solution are diluted with the same solution.

2. Preparation of BCA working solution: mixing 50 parts of BCA Reagent with 1 part of copper sulfate solution (50:1, BCA Reagent: copper sulfate solution). BCA working solution can be stored at room temperature and used within 24h. Add 200 µL of the BCA working solution to each well. It is recommended to prepare as required to avoid waste. 

3. Detection: add 200 µL of the BCA working solution to each well and mix plate thoroughly on a plate shaker for 30 seconds; After cover plate and incubate at 37°C for 30 minutes, the standard curve No. 0 was used as a reference to measure the absorbance at or near 562 nm on a plate reader. (Note: it can also be reacted at room temperature for 2 h or 60℃ for 30 min. If the protein concentration is low, it is recommended to react at 60℃)

4. Calculation: the standard curve was drawn with gradient protein content (μg/mL) as abscissa and absorption value as ordinate. According to the absorbance value of the test-sample, the protein concentration (μg/mL) of the test-sample in the corresponding well can be found on the standard curve, and then multiplied by the dilution factor of the sample is the actual protein concentration of the test-sample. 

Note

1. Determination of protein concentration by BCA method is greatly affected by temperature and time, and the absorbance value will change with the extension of time or the increase of temperature. If the time and temperature of color reaction cannot be accurately controlled, it is recommended to make a standard curve for each determination.

2. When preparing protein standard storage solution, it is necessary to ensure sufficient dissolution. Dilution 10 times gradient dilution is recommended when preparing protein standard working solution. Do not dilute 50 times at a time to avoid error.

3. In order to guarantee quantification of protein, it is better to choose the same buffer solution for sample extraction and protein standard dilution to ensure the same detection conditions. If the buffer has a high background value, other methods are recommended.

4. Please wear experimental suits and disposable gloves when operation.

Reference

• Cortez Karoll, J., et al.Infections Caused by Scedosporium spp.Clinical Microbiology Reviews21,157-197(2008).PMID 18202441IF 36.8 / PMID:18202441

• Wang, Z., et al.FUT2-dependent fucosylation of HYOU1 protects intestinal stem cells against inflammatory injury by regulating unfolded protein response.Redox Biology60,102618(2023).PMID 36724577IF 11.4 / PMID:36724577

• Zhang, L., et al.Embryonic Exposure to UV-328 Impairs the Cell Cycle in Zebrafish (Danio rerio) by Inhibiting the p38 MAPK/p53/Gadd45a Signaling Pathway.Environmental Science & Technology(2023).PMID 37384941IF 11.4 / PMID:37384941

• Zhang, S., et al.A novel PHD2 inhibitor acteoside from Cistanche tubulosa induces skeletal muscle mitophagy to improve cancer-related fatigue.Biomedicine & Pharmacotherapy150,113004(2022).PMID 35658245IF 7.5 / PMID:35658245

• Xia, Q.-s., et al.Berberine reduces hepatic ceramide levels to improve insulin resistance in HFD-fed mice by inhibiting HIF-2α.Biomedicine & Pharmacotherapy150,112955(2022).PMID 35429745IF 7.5 / PMID:35429745

• Shi, R., et al.CircESRP1 enhances metastasis and epithelial–mesenchymal transition in endometrial cancer via the miR-874-3p/CPEB4 axis.Journal of Translational Medicine20,139(2022).PMID 35317822IF 7.4 / PMID:35317822

• Dong, L., et al.Anti-inflammatory effect of Rhein on ulcerative colitis via inhibiting PI3K/Akt/mTOR signaling pathway and regulating gut microbiota.Phytotherapy Research36,2081-2094(2022).PMID 35229916IF 7.2 / PMID:35229916

• Chen, Y., et al.Inhibiting mitochondrial inflammation through Drp1/HK1/NLRP3 pathway: A mechanism of alpinetin attenuated aging-associated cognitive impairment.Phytotherapy Research37,2454-2471(2023).PMID 36772986IF 7.2 / PMID:36772986

• Huang, P., W. Jin, and S. XuOptimization of smashing tissue and ultrasonic extraction of tanshinones and their neuroprotective effect on cerebral ischemia/reperfusion injury by inhibiting parthanatos.Food Funct13,9658-9673(2022).PMID 36040108IF 6.1 / PMID:36040108

• Yu, W., et al.Chemical composition and anti-inflammatory activities of Castanopsis honey.Food Funct14,250-261(2023).PMID 36484340IF 6.1 / PMID:36484340

• Xin, J., et al.Spatial transcriptomics analysis of zone-dependent hepatic ischemia-reperfusion injury murine model.Communications Biology6,194(2023).PMID 36804628IF 5.9 / PMID:36804628

• Xie, Y., et al.Gamma-Muricholic Acid Inhibits Nonalcoholic Steatohepatitis: Abolishment of Steatosis-Dependent Peroxidative Impairment by FXR/SHP/LXRα/FASN Signaling.Nutrients15,(2023).PMID 36904254IF 5.9 / PMID:36904254

• Zhuang, H., et al.Indole-3-aldehyde alleviates chondrocytes inflammation through the AhR-NF-κB signalling pathway.International Immunopharmacology113,109314(2022).PMID 36252481IF 5.6 / PMID:36252481

• Ren, X., et al.Ceria Nanoparticles Alleviated Osteoarthritis through Attenuating Senescence and Senescence-Associated Secretory Phenotype in Synoviocytes.Int J Mol Sci24,(2023).PMID 36902483IF 5.6 / PMID:36902483

• Lv, S., et al.Anxiolytic effect of YangshenDingzhi granules: Integrated network pharmacology and hippocampal metabolomics.Frontiers in Pharmacology13,(2022).PMID 36386232IF 5.6 / PMID:36386232

• Ren, X., et al.Gsmtx4 Alleviated Osteoarthritis through Piezo1/Calcineurin/NFAT1 Signaling Axis under Excessive Mechanical Strain.Int J Mol Sci24,(2023).PMID 36835440IF 5.6 / PMID:36835440

• Xie, Y., et al.Polygoni Multiflori Radix Praeparata and Acori Tatarinowii Rhizoma ameliorate scopolamine-induced cognitive impairment by regulating the cholinergic and synaptic associated proteins.Journal of Ethnopharmacology311,116400(2023).PMID 37003402IF 5.4 / PMID:37003402

• Jiang, Y., et al.Integrated strategy of network pharmacology, molecular docking, HPLC-DAD and mice model for exploring active ingredients and pharmacological mechanisms of Penthorum chinense Pursh against alcoholic liver injury.Journal of Ethnopharmacology298,115589(2022).PMID 35926779IF 5.4 / PMID:35926779

• Qian, Y., et al.Seeds of Ginkgo biloba L. inhibit oxidative stress and inflammation induced by cigarette smoke in COPD rats through the Nrf2 pathway.Journal of Ethnopharmacology301,115758(2023).PMID 36167232IF 5.4 / PMID:36167232

• Mei, Y., et al.The effect of gastric bypass surgery on cognitive function of Alzheimer’s disease and the role of GLP1-SGLT1 pathway.Experimental Neurology363,114377(2023).PMID 36893833IF 5.3 / PMID:36893833

• Niu, H., et al.Effect of Lactobacillus rhamnosus MN-431 Producing Indole Derivatives on Complementary Feeding-Induced Diarrhea Rat Pups Through the Enhancement of the Intestinal Barrier Function.Molecular Nutrition & Food Research66,2100619(2022).PMID 34806832IF 5.2 / PMID:34806832

• Jiang, H., et al.Yinchen Linggui Zhugan decoction ameliorates high fat diet-induced nonalcoholic fatty liver disease by modulation of SIRT1/Nrf2 signaling pathway and gut microbiota.Frontiers in Microbiology13,(2022).PMID 36578580IF 5.2 / PMID:36578580

• Fan, X., et al.Light intensity alters the effects of light-induced circadian disruption on glucose and lipid metabolism in mice.American Journal of Physiology-Endocrinology and Metabolism322,E1-E9(2021).PMID 34719945IF 5.1 / PMID:34719945

• Li, Z.-R., et al.Sleep deprivation leads to further impairment of hippocampal synaptic plasticity by suppressing melatonin secretion in the pineal gland of chronically unpredictable stress rats.European Journal of Pharmacology930,175149(2022).PMID 35878808IF 5 / PMID:35878808

• Song, X., et al.Cochlear resident macrophage mediates development of ribbon synapses via CX3CR1/CX3CL1 axis.Frontiers in Molecular Neuroscience15,(2022).PMID 36518186IF 4.8 / PMID:36518186

• Xu, D., et al.Activation of the PACAP/PAC1 Signaling Pathway Accelerates the Repair of Impaired Spatial Memory Caused by an Ultradian Light Cycle.ASN Neuro15,17590914231169140(2023).PMID 37071544IF 4.7 / PMID:37071544

• Yao, Z., et al.The expression of IGFBP-5 in the reproductive axis and effect on the onset of puberty in female rats.Reproductive Biology and Endocrinology20,100(2022).PMID 35821045IF 4.4 / PMID:35821045

• Wei, W., et al.Network pharmacology reveals that Berberine may function against Alzheimer’s disease via the AKT signaling pathway.Frontiers in Neuroscience17,(2023).PMID 37214397IF 4.3 / PMID:37214397

• Talifu, Z., et al.The Overexpression of Insulin-Like Growth Factor-1 and Neurotrophin-3 Promote Functional Recovery and Alleviate Spasticity After Spinal Cord Injury.Frontiers in Neuroscience16,(2022).PMID 35573286IF 4.3 / PMID:35573286

• Lei, X., et al.Activation of HCN channels caused by elevated cAMP levels in periaqueductal gray promotes bone cancer pain.Neurochemistry International162,105437(2023).PMID 36336089IF 4.2 / PMID:36336089

• Fei, W.-t., et al.Immunomodulatory Effects of <i>Lepidium meyenii</i> Walp. Polysaccharides on an Immunosuppression Model Induced by Cyclophosphamide.Journal of Immunology Research2022,1210890(2022).PMID 35832646IF 4.1 / PMID:35832646

• Si, Y., et al.Phellodendronoside A Exerts Anticancer Effects Depending on Inducing Apoptosis Through ROS/Nrf2/Notch Pathway and Modulating Metabolite Profiles in Hepatocellular Carcinoma.Journal of Hepatocellular Carcinoma10,935-948(2023).PMID 37361906IF 4.1 / PMID:37361906

• Xiao, J., et al.TM9SF1 knockdown decreases inflammation by enhancing autophagy in a mouse model of acute lung injury.Heliyon8,e12092(2022).PMID 36561687IF 4 / PMID:36561687

• Xie, B., et al.DNA methylation influences the CTCF-modulated transcription of RASSF1A in lung cancer cells.Cell Biology International46,1900-1914(2022).PMID 35989484IF 3.9 / PMID:35989484

• Lv, S., et al.Ginsenoside Rg3 induces apoptosis and inhibits proliferation by down-regulating TIGAR in rats with gastric precancerous lesions.BMC Complementary Medicine and Therapies22,188(2022).PMID 35840932IF 3.9 / PMID:35840932

• Wang, Z., et al.NLRP3 inflammasome deficiency attenuates cerebral ischemia-reperfusion injury by inhibiting ferroptosis.Brain Research Bulletin193,37-46(2023).PMID 36435361IF 3.8 / PMID:36435361

• Xie, Y., et al.Profile analysis and functional modeling identify circular RNAs in nonalcoholic fatty liver disease as regulators of hepatic lipid metabolism.Frontiers in Genetics13,(2022).PMID 36186461IF 3.7 / PMID:36186461

• Li, X., et al.Identification of the Prognostic Biomarkers CBX6 and CBX7 in Bladder Cancer.Diagnostics (Basel)13,(2023).PMID 37189494IF 3.6 / PMID:37189494

• Fan, K., et al.ERBB2 S310F mutation independently activates PI3K/AKT and MAPK pathways through homodimers to contribute gallbladder carcinoma growth.Medical Oncology39,64(2022).PMID 35477796IF 3.4 / PMID:35477796

• Jiang, G., et al.Autophagy: A New Mechanism for Esketamine as a Depression Therapeutic.Neuroscience498,214-223(2022).PMID 35597333IF 3.3 / PMID:35597333

• Li, J., et al.Cadherin-11 promotes the mechanical strength of engineered elastic cartilage by enhancing extracellular matrix synthesis and microstructure.Journal of Tissue Engineering and Regenerative Medicine16,188-199(2022).PMID 34837334IF 3.3 / PMID:34837334

• Xie, Z., et al.Inhibition of Ferroptosis Attenuates Glutamate Excitotoxicity and Nuclear Autophagy in a CLP Septic Mouse Model.Shock57,694-702(2022).PMID 35066511IF 3.1 / PMID:35066511

• Li, P., et al.New Target of Oxidative Stress Regulation in Cochleae: Alternative Splicing of the p62/Sqstm1 Gene.Journal of Molecular Neuroscience72,830-840(2022).PMID 35048235IF 3.1 / PMID:35048235

• Yu, J., et al.Quercetin Ameliorates Lipopolysaccharide-Induced Duodenal Inflammation through Modulating Autophagy, Programmed Cell Death and Intestinal Mucosal Barrier Function in Chicken Embryos.Animals (Basel)12,(2022).PMID 36552443IF 3 / PMID:36552443

• Jia, S., et al.Anti-Neuroinflammatory Activity of New Naturally Occurring Benzylated Hydroxyacetophenone Analogs from the Endophytic Fungus Alternaria sp.Chemistry & Biodiversity19,e202200751(2022).PMID 36082622IF 2.9 /  PMID:36082622 

• Li, H., et al.Knockdown of lncRNA Meg3 delays the onset of puberty in female rats.Theriogenology207,72-81(2023).PMID 37269598IF 2.8 / PMID:37269598

• Xiong, R., et al.Oral Hydroxychloroquine Mitigates Lipopolysaccharide-induced Lung Injury by Inhibiting Pyroptosis in Mice.Curr Mol Pharmacol16,362-373(2023).PMID 35996240IF 2.7 / PMID:35996240

• Bao, W.-A., et al.Baicalin Ameliorates Radiation-Induced Lung Injury by Inhibiting the CysLTs/CysLT1 Signaling Pathway.Evidence-Based Complementary and Alternative Medicine2022,2765354(2022).PMID 35783527IF 2.6 / PMID:35783527

• Chen, S., et al.BHLHE41 Overexpression Alleviates the Malignant Behavior of Colon Cancer Cells Induced by Hypoxia via Modulating HIF-1<i>α</i>/EMT Pathway.Gastroenterology Research and Practice2022,6972331(2022).PMID 35615737IF 2 / PMID:35615737

For Research Use Only!