Protein fractions were also analyzed by SDS-PAGE denaturing gels followed by staining with SimplyBlue SafeStain solution (Invitrogen) or Western blotting as described (15 (link)). For Western blot analysis, the following primary antibodies were used: BRCA1 (C-20; Santa Cruz Biotechnology, sc-642), ubiquitin-pAb (Enzo Life Sciences, ADI-SPA-200), p53 (DO-1; Santa Cruz Biotechnology, sc-126), K63-linkage specific polyubiquitin (D7A11; Cell Signaling, #5621) and β-actin (Sigma-Aldrich, A5441). Western blot quantification and densitometry measurements were performed using Image Lab™ Software (Bio-Rad). The band intensities were selected using the volume tool. Local subtraction and linear regression methods were implemented to eliminate the local background values and to quantify the band intensities.
Brca1 c20
Manufactured by Santa Cruz Biotechnology
Sourced in United States
BRCA1 (C20) is a primary antibody targeting the BRCA1 protein. BRCA1 is a tumor suppressor gene that plays a critical role in DNA repair and cell cycle regulation. The BRCA1 (C20) antibody is a tool for researchers studying the BRCA1 protein and its functions.
Automatically generated - may contain errors
Lab products found in correlation
Simplyblue safe stain solution, by Thermo Fisher Scientific (1 mentions)
K63 linkage specific polyubiquitin d7a11, by Cell Signaling Technology (1 mentions)
P53 do 1, by Santa Cruz Biotechnology (1 mentions)
β actin, by Merck Group (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
Rap80, by Fortis Life Sciences (1 mentions)
H3k9me2, by Merck Group (1 mentions)
Rnf168, by Merck Group (1 mentions)
γh2ax jbw301, by Merck Group (1 mentions)
Benzonase nuclease, by Merck Group (1 mentions)
Jbw301, by Merck Group (1 mentions)
Sqstm1 p62 d5e2, by Cell Signaling Technology (1 mentions)
β actin, by Abcam (1 mentions)
3 protocols using brca1 c20
1
SDS-PAGE and Western Blot Analysis
2
Antibody-based Analysis of DNA Damage Response
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The antibodies used against FANCJ (Bethyl Laboratories, Montgomery, TX, USA), BARD1 (BL518; Bethyl Laboratories), BRCA1 (C20; Santa Cruz Biotechnology, Santa Cruz, CA, USA), KAP‐pS824 (Bethyl Laboratories), RAP80, (Bethyl Laboratories), RNF168 (ABE367; Millipore, Darmstadt, Germany), and RAD51 (BioAcademia, Osaka, Japan) were rabbit polyclonal antibodies. The antibodies against CtIP (Active Motif, Carlsbad, CA, USA), conjugated ubiquitin (FK2; Nippon Bio‐Test Laboratories, Tokyo, Japan), HP1α (Millipore), HP1β (1MOD‐1A9; Millipore), HP1γ (2MOD‐1G6; Millipore), H3K9me2 (CMA307; Millipore), γH2AX (JBW301; Millipore), and α‐ and β‐tubulin (DMIA+BMIB; Neomarkers, Fremont, CA, USA) were mouse mAbs.
3
Immunoblotting of Nuclear Proteins
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Cell lysates were prepared with 0.5% NP-40 buffer (50 mM Tris–HCl [pH 7.5], 0.5% Nonidet P-40, 150 mM NaCl, 50 mM NaF, 1 mM dithiothreitol, 1 mM NaVO3, 1 mM PMSF, 2 µg/ml aprotinin, 2 µg/ml leupeptin, 10 µg/ml trypsin inhibitor, and 150 µg/ml benzamidine) and subjected to immunoblotting, as previously described [15 (link), 16 (link)]. Benzonase nuclease (125 U/ml; Millipore, MA, USA) and 2 mM MgCl2 were supplemented for detection of nuclear proteins. The relative protein expression levels of Trop-2 were measured using a densitometer and normalized to tubulin. The following antibodies were used: rabbit monoclonal antibody against Trop-2 (Abcam, Cambridge, UK), AKT (Cell Signaling Tech, MA, USA), SQSTM1/p62 (D5E2; Cell Signaling Tech), rabbit polyclonal antibody against ERα (Millipore), BRCA1 (C20; Santa Cruz, CA, USA), HER2 (Cell Signaling Tech), pAKT (phospho-Ser473 AKT; Cell Signaling Tech), TFEB (Cell Signaling Tech), pRSK (Pan phospho-Ser221, Ser227, Ser218, Ser232; Invitrogen), LC3 (2775; Cell Signaling Tech), mouse monoclonal antibody against γH2AX (JBW301, Millipore), RSK1 (A-10; Santa Cruz), RSK2 (E-1; Santa Cruz), β-actin (6276; Abcam), and α-tubulin (DM1A; Richard-Allan Scientific, MI, USA).
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