MA1-23164

antibody from Invitrogen Antibodies

Targeting: BRCA1 BRCC1, FANCS, PPP1R53, RNF53

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-23164

Provider

Invitrogen Antibodies

Product name

BRCA1 Monoclonal Antibody (6B4)

Antibody type

Monoclonal

Antigen

Other

Description

The predicted molecular weight is 220 kDa. A suggested positive control for this product is HBL100.

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

6B4

Vial size

100 µL

Concentration

2.75 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Human Papillomaviruses Preferentially Recruit DNA Repair Factors to Viral Genomes for Rapid Repair and Amplification.

Mehta K, Laimins L
mBio 2018 Feb 13;9(1)

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Analysis of alternative lengthening of telomere markers in BRCA1 defective cells.

Kargaran PK, Yasaei H, Anjomani-Virmouni S, Mangiapane G, Slijepcevic P
Genes, chromosomes & cancer 2016 Nov;55(11):864-76

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of BRCA1 in MCF-7 and MEF (mouse embryonic fibroblast) cell lysate. Samples were resolved on a 5-15% gradient gel and probed with a BRCA1 monoclonal antibody (Product # MA1-23164).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of BRCA1 was performed by separating 30 and 50 µg of whole cell extracts by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a BRCA1 Monoclonal Antibody (6B4) (Product # MA1-23164) at a dilution of 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of BRCA1 was performed by separating 30 and 50 µg of whole cell extracts by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a BRCA1 Monoclonal Antibody (6B4) (Product # MA1-23164) at a dilution of 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

BRCA1 Polyclonal Antibody [6B4] BRCA1 Monoclonal Antibody (6B4) (Product # MA1-23164) was used at 1:1,000 dilution for western blot assay of lysates from cells transfected with control or BRCA1-specific siRNA. Lysates were prepared at the indicated times following transfection. RAD50 Monoclonal Antibody (13B3) (Product # MA1-23269) was used as a loading control.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

BRCA1 antibody 6B4 (Product # MA1-23164) was used for immunofluorescent staining of BRCA1 nuclear foci induced by ionizing radiation. IR-treated (2 hr /4 gray IR) U2OS cells were pre-extracted with CSK buffer on ice for 4 min before fixation with 4% PFA in room temperature, and then subjected to immunostaining. DAPI was used to counterstain nucleus. 6B4 was used at 1:400 dilution. Secondary antibody (Alexa Fluor-488) used for detection of primary antibody (BRCA1 antibody 6B4).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis using BRCA1 Monoclonal Antibody (6B4) (Product # MA1-23164).

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

BRCA1 antibody 6B4 (Product # MA1-23164) and BRCA1 antibody 17F8 (Product # MA1-23160) were used for ChIP assay. The 6B4 and 17F8 mixture (3 microgram each), or normal mouse IgG (6 microgram) were incubated with HeLa chromatin extract (100 microgram each) in the ChIP assay. Enrichment of genomic DNA on a BRCA1 target gene promoter (HMGA2) was validated by a Q-PCR assay.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

BRCA1 antibody 6B4 (Product # MA1-23164) and BRCA1 antibody 17F8 was used for IP-Western Blot assay. 6B4 alone (4 µg), 17F8 alone (4 µg), 6B4 plus 17F8 (2 µg each), and mouse control normal IgG were used in an immunoprecipitation assay with MCF7 cell extract. Immunoprecipitated BRCA1 was detected in Western Blot using BRCA1 antibody 6B4 at 1:1,000 dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Double immunofluorescence analysis of APBs, BRCA1, ATRX, BLM in the human and mouse cells: (A and B) Total number of ALT-associated PML bodies (APBs) in the human and mouse cells. There were significantly higher levels of APBs in the U2OS ALT-positive human cells and the Brca1 defective mESC 408; however, the levels of APBs associated with telomeres were not significantly different in the mESC 408 but was much higher in the U2OS cells. (C and D) Reduced BLM foci were observed in the U2OS ALT-positive human cells but not in the ALT negative human or the ALT-positive mouse cells. However, BLM colocalizations with telomeres were significantly higher in the U2OS and Brca1 wt mESC E14. (E) Colocalization of BRCA1 with ATRX, and (F) BRCA1 with BLM, reveal differences between human and mouse cells. A minimum of 50 cell nuclei was analyzed in two-independent experiments in all the samples shown above. The error bars represent min/max in the whisker box plot and the gray dotted line indicates mean in the scatter graphs. t -Test used for statistical analysis with alpha set at 0.05.