Anti-BRMS1 antibody [EPR7202]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRMS1 antibody [EPR7202] (AB134968)

Immunohistochemical analysis using ab134968 showing positive staining in Ovarian carcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BRMS1 antibody [EPR7202] (AB134968)

Immunofluorescence staining of HepG2 cells with purified ab134968 at a working dilution of 1/1000 counter-stained with DAPI. The secondary antibody was an Alexa Fluor^® 488 conjugated goat anti-rabbit (ab150077) used at a dilution of 1/1000. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control PBS was used instead of the primary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRMS1 antibody [EPR7202] (AB134968)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling BRMS1 with ab134968 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRMS1 antibody [EPR7202] (AB134968)

Imunohistochemical analysis of paraffin-embedded Human placenta tissue labelling BRMS1 with ab134968 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRMS1 antibody [EPR7202] (AB134968)

Immunohistochemical analysis using ab134968 showing positive staining in Breast carcinoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-BRMS1 antibody [EPR7202] (AB134968)

BRMS1 was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg with 134968 at 1/50 dilution (2μg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg

Lane 2 : ab134968 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab134968 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-BRMS1 antibody [EPR7202] (ab134968)

Predicted band size: 28 kDa

Observed band size: 28 kDa

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• WB

Unknown

Western blot - Anti-BRMS1 antibody [EPR7202] (AB134968)

All lanes:

Western blot - Anti-BRMS1 antibody [EPR7202] (ab134968) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Lane 4:

Human colon cancer lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 28 kDa

Observed band size: 28 kDa

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• WB

CiteAb

Western blot - Anti-BRMS1 antibody [EPR7202] (AB134968)

BRMS1 western blot using anti-BRMS1 antibody [EPR7202] ab134968. Publication image and figure legend from Krämer, D., Schön, M., et al., 2015, Cell Death Dis, PubMed 25675294.

ab134968 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab134968 please see the product overview.

BRMS1 mediates the destabilization of p300 and CBP upon iASPP removal and cisplatin treatment. (a) BRMS1 knockdown can rescue p300 and CBP levels in cisplatin-treated, iASPP-depleted cells. BRMS1 was transiently knocked down with siRNA in cells that expressed control or iASPP shRNA. Scrambled siRNA was transfected in parallel for control. 48 h after knockdown, the cells were either left untreated or incubated with cisplatin (24 h, 20 μM) followed by immunoblot analysis. (b) iASPP overexpression abrogates the interaction of BRMS1 and CBP in cisplatin-treated cells. HCT116 cells were transiently transfected with a BRMS1-GFP expression construct, alone or in combination with the iASPP-V5 expression plasmid. 24 h after transfection, the cells were treated with 20 μM cisplatin for 16 h and 20 μM MG132 for the last 4 h. Co-immunoprecipitation was performed using either an antibody for CBP or an isotype-matched control antibody (IgG). (c) Proposed mechanism of iASPP-mediated apoptosis. iASPP inhibits BRMS1-dependent proteasomal degradation of p300/CBP, thereby enhancing the induction of pro-apoptotic TAp73 target genes

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