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Pierce s bca protein assay reagent kit

Manufactured by Thermo Fisher Scientific
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Sourced in United States

The Pierce's BCA Protein Assay Reagent Kit is a colorimetric detection and quantitation assay used to determine the total protein concentration in a sample. The kit utilizes the bicinchoninic acid (BCA) method to measure the reduction of copper ions by proteins in an alkaline environment, resulting in a purple-colored reaction that can be measured spectrophotometrically.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) Antibiotic antimycotic, by Merck Group (4 mentions) Dharmafect1 t 2001 03, by Thermo Fisher Scientific (3 mentions) Dharmafect 1, by Thermo Fisher Scientific (2 mentions) Hop62, by American Type Culture Collection (1 mentions) H2009, by American Type Culture Collection (1 mentions) Antibiotics antimycotics, by Merck Group (1 mentions) Sk n be 2, by American Type Culture Collection (1 mentions) Imr 90, by American Type Culture Collection (1 mentions) Ceb lysis buffer, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Pierce BCA reagents Pierce BCA assay BCA protein reagent Pierce BCA kit BCA protein assay BCA assay reagent BCA protein kit BCA reagent kit Pierce BCA Protein assay reagent

5 protocols using pierce s bca protein assay reagent kit

1

Culturing and Transfecting Non-Small Cell Lung Cancer Cell Lines

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Human non-small cell lung carcinoma cell line A549 and H358 were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) and cultured in RPMI medium supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) and 1% antibiotic/antimycotic (Sigma, St Louis, MO, USA). The cell lines were routinely subcultured every 3–4 days. All siRNA transfections were performed using Dharmafect1 #T-2001-03 (Thermo Fisher Scientific Inc, Pittsburgh, PA, USA) as per manufacturer’s protocol. After total 72 hrs of transfection cells were harvested in CHAPS lysis buffer (1% CHAPS detergent, 150mM NaCl, 50mM Tris pH 7, 5mM EDTA). Protein was quantitated by using Pierce's BCA Protein Assay Reagent Kit (# 23227) from Pierce Biotechnology, Rockford, IL, USA as per manufacturer’s protocol.
Shah P.P., Lockwood W.W., Saurabh K., Kurlawala Z., Shannon S.P., Waigel S., Zacharias W, & Beverly L.J. (2014). Ubiquilin1 Represses Migration and Epithelial to Mesenchymal Transition of Human Non-small Cell Lung Cancer Cells. Oncogene, 34(13), 1709-1717.
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2

Culturing and Transfecting Lung Cancer Cell Lines

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Human lung adenocarcinoma cell lines A549, HOP62, H23, and H2009 were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) and cultured in RPMI medium supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) and 1% antibiotic/antimycotic (Sigma, St Louis, MO, USA). Human Embryonic Kidney cells (293T; ATCC® CRL3216TM) were cultured in DMEM medium supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) and 1% antibiotic/antimycotic (Sigma, St Louis, MO, USA). The cell lines were routinely subcultured every 3–5 days. All siRNA transfections were performed using Dharmafect1 # T-2001-03 (Thermo Fisher Scientific Inc, Pittsburgh, PA, USA) as per the manufacturer’s protocol. After a total of 72 h of transfection cells were harvested in CEB lysis buffer # FNN0011 (Invitrogen, Life Technologies, Grand Island, NY, USA, 14072). Protein was quantitated by using Pierce’s BCA Protein Assay Reagent Kit (#23227) from Pierce Biotechnology, Rockford, IL, USA as per the manufacturer’s protocol.
Shah P.P, & Beverly L.J. (2023). UBQLN Family Members Regulate MYC in Lung Adenocarcinoma Cells. Cancers, 15(13), 3389.
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3

IGF1 Signaling in Lung Adenocarcinoma

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Human non-small cell lung adenocarcinoma cell lines A549 were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) and cultured in RPMI medium supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) and 1% antibiotics/antimycotics (Sigma, St Louis, MO, USA). The 3 cell lines were routinely sub-cultured every 3–4 days. All siRNA transfections were performed using Dharmafect1 #T-2001-03 (Thermo Fisher Scientific Inc, Pittsburgh, PA, USA) as per manufacturer’s protocol. After total 48 hours of transfection, cells were serum starved for 12 hours. After which IGF1 (50ng/ml) was added to stimulate the cells. For protein stability studies, Cycloheximide (20uM), and for testing degradation pathways, Monensin (10uM) were added an hour prior to stimulating with IGF1. At the end of stimulation for various time points, cells were harvested in CHAPS lysis buffer (1% CHAPS detergent, 150mM NaCl, 50mM Tris pH 7, 5mM EDTA) supplemented with protease and phosphatase inhibitors. Protein was quantified by using Pierce's BCA Protein Assay Reagent Kit (#23227) from Pierce Biotechnology, Rockford, IL, USA as per manufacturer’s protocol.
Kurlawala Z., Dunaway R., Shah P.P., Gosney J.A., Siskind L.J., Ceresa B.P, & Beverly L.J. (2017). Regulation of Insulin-like Growth Factor Receptors by Ubiquilin1. The Biochemical journal, 474(24), 4105-4118.
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4

Comparative Analysis of Cell Line Responses

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Human lung adenocarcinoma cell line A549, neuroblastoma cell line SK-N-BE(2) and IMR90 fetal lung fibroblasts were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) and cultured in RPMI medium supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) and 1% antibiotic/antimycotic (Sigma, St Louis, MO, USA). The cell lines were routinely tested for mycoplasma and subcultured every 3–4 days. All siRNA transfections were performed using Dharmafect1 #T-2001-03 (Thermo Fisher Scientific Inc, Pittsburgh, PA, USA) as per manufacturer's protocol. After total 72 hrs of transfection cells were harvested in CHAPS lysis buffer (1% CHAPS detergent, 150mM NaCl, 50mM Tris pH 7, 5mM EDTA). Protein was quantitated by using Pierce's BCA Protein Assay Reagent Kit (# 23227) from Pierce Biotechnology, Rockford, IL, USA as per manufacturer's protocol.
Shah P.P., Saurabh K., Kurlawala Z., Vega A.A., Siskind L.J, & Beverly L.J. (2022). Towards a molecular understanding of the overlapping and distinct roles of UBQLN1 and UBQLN2 in lung cancer progression and metastasis. Neoplasia (New York, N.Y.), 25, 1-8.
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5

Lung Cancer Cell Line Cultivation and siRNA Transfection

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Human adenocarcinoma cell lines A549, H358 and human immortalized small airway epithelial cell line (HPLD-1) were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) and cultured in RPMI medium supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) and 1% antibiotic/antimycotic (Sigma, St Louis, MO, USA). The cell lines were routinely subcultured every 3–5 days. All siRNA transfections were performed using Dharmafect1 # T-2001-03 (Thermo Fisher Scientific Inc, Pittsburgh, PA, USA) as per manufacturer's protocol. After total 72 hrs of transfection cells were harvested in CEB lysis buffer # FNN0011 (Invitrogen, Life technologies, Grand Island, NY, 14072). Protein was quantitated by using Pierce's BCA Protein Assay Reagent Kit (# 23227) from Pierce Biotechnology, Rockford, IL, USA as per manufacturer's protocol (for more detail regarding siRNA sequences used for study, see Supplemental Methods).
Shah P.P, & Beverly L.J. (2015). Regulation of VCP/p97 demonstrates the critical balance between cell death and epithelial-mesenchymal transition (EMT) downstream of ER stress. Oncotarget, 6(19), 17725-17737.
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