Product Name
BRCC36 Recombinant Rabbit Monoclonal Antibody [PSH05-10] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human BRCC36 aa 1-316 / 316.
Species Reactivity
Human, Mouse, Rat, Green monkey
Target Molecular Weight
Predicted band size: 36 kDa
Positive Control
PC-3M cell lysate, PC-3 cell lysate, HeLa cell lysate, MCF7 cell lysate, HEK-293 cell lysate, SK-Br-3 cell lysate, HT-29 cell lysate, COS-1 cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, PC-12 cell lysate, L6 cell lysate, C6 cell lysate, human heart tissue, human prostate cancer tissue, human kidney tissue, mouse kidney tissue, rat kidney tissue, PC-3M, Neuro-2a, C6.
Storage Instructions
Store at 2-8℃. Avoid freeze.
Purification Method
Protein A affinity purified.
Function
Lys-63-specific deubiquitinase BRCC36 is an enzyme that in humans is encoded by the BRCC3 gene. This gene encodes a subunit of the BRCA1-BRCA2-containing complex (BRCC), which is an E3 ubiquitin ligase. This protein is also thought to be involved in the cellular response to ionizing radiation and progression through the G2/M checkpoint. Alternative splicing results in multiple transcript variants. BRCC36, the protein product of the BRCC3 gene, is a deubiquitinating enzyme and a core component of the deubiquitin complex BRCA1-A. BRCA1, as distinct from BRCA1-A, is employed in the repair of chromosomal damage with an important role in the error-free homologous recombinational (HR) repair of DNA double-strand breaks. Sequestration of BRCA1 away from the DNA damage site suppresses homologous recombination and redirects the cell in the direction of repair by the process of non-homologous end joining (NHEJ). The role of BRCA1-A appears to be to bind BRCA1 with high affinity and withdraw it away from the site of DNA damage to the periphery where it remains sequestered, thus promoting DNA repair by NHEJ in preference to HR.
Background References
1. Cheng Q et al. BRCC36 functions noncatalytically to promote antiviral response by maintaining STAT1 protein stability. Eur J Immunol. 2021 Feb
2. Wang H et al. BRCC36 Deubiquitinates HMGCR to Regulate the Interplay Between Ferroptosis and Pyroptosis. Adv Sci (Weinh). 2024 Jan
Subcellular Location
Nucleus, Cytoplasm, cytoskeleton, spindle pole.
Synonyms
BRCA1-A complex subunit BRCC36 antibody
BRCA1/BRCA2 containing complex subunit 3 antibody
BRCA1/BRCA2 containing complex subunit 36 antibody
BRCA1/BRCA2-containing complex subunit 3 antibody
BRCA1/BRCA2-containing complex subunit 36 antibody
Brcc3 antibody
BRCC3_HUMAN antibody
BRCC36 antibody
BRISC complex subunit BRCC36 antibody
C6.1A antibody
Expand
BRCA1-A complex subunit BRCC36 antibody
BRCA1/BRCA2 containing complex subunit 3 antibody
BRCA1/BRCA2 containing complex subunit 36 antibody
BRCA1/BRCA2-containing complex subunit 3 antibody
BRCA1/BRCA2-containing complex subunit 36 antibody
Brcc3 antibody
BRCC3_HUMAN antibody
BRCC36 antibody
BRISC complex subunit BRCC36 antibody
C6.1A antibody
Chromosome X open reading frame 53 antibody
CXorf53 antibody
Lys-63-specific deubiquitinase BRCC36 antibody
OTTHUMP00000061450 antibody
Collapse
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Western blot analysis of BRCC36 on different lysates with Rabbit anti-BRCC36 antibody (HA722226) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.
Lane 1: PC-3M cell lysate
Lane 2: PC-3 cell lysate
Lane 3: HeLa cell lysate
Lane 4: MCF7 cell lysate
Lane 5: HEK-293 cell lysate
Lane 6: SK-Br-3 cell lysate
Lane 7: HT-29 cell lysate
Lane 8: COS-1 cell lysate
Lane 9: Neuro-2a cell lysate
Lane 10: NIH/3T3 cell lysate
Lane 11: C2C12 cell lysate
Lane 12: PC-12 cell lysate
Lane 13: L6 cell lysate
Lane 14: C6 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 36 kDa
Observed band size: 33/36 kDa
Exposure time: 25 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722226) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human heart tissue with Rabbit anti-BRCC36 antibody (HA722226) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722226) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue with Rabbit anti-BRCC36 antibody (HA722226) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722226) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-BRCC36 antibody (HA722226) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722226) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-BRCC36 antibody (HA722226) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722226) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-BRCC36 antibody (HA722226) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722226) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of PC-3M cells labeling BRCC36 with Rabbit anti-BRCC36 antibody (HA722226) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BRCC36 antibody (HA722226) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of Neuro-2a cells labeling BRCC36 with Rabbit anti-BRCC36 antibody (HA722226) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BRCC36 antibody (HA722226) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of C6 cells labeling BRCC36 with Rabbit anti-BRCC36 antibody (HA722226) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BRCC36 antibody (HA722226) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of Neuro-2a cells labeling BRCC36.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722226, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
-
This data was developed using HA722226, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of C6 cells labeling BRCC36.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722226, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"