MA5-42558

antibody from Invitrogen Antibodies

Targeting: CENPB

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-42558

Provider

Invitrogen Antibodies

Product name

CENPB Monoclonal Antibody (A5F5)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

Positive Control: Hela cell lysate, A549 cell lysate, MCF-7 cell lysate, A549, human kidney tissue, human skin tissue.

Antibody clone number

A5F5

Concentration

2 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of CENPB on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. CENPB Monoclonal Antibody (Product # MA5-42558) at 1:500 was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:20,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Hela cell lysate. Lane 2: A549 cell lysate. Lane 3: MCF-7 cell lysate. Predicted band size: 65 kDa. Observed band size: 80 kDa.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

CENPB was immunoprecipitated from 0.4 mg Hela whole cell lysates with CENPB Monoclonal Antibody (Product # MA5-42558) at 2 μg/mL. Western blot was performed from the immunoprecipitate using CENPB Monoclonal Antibody (Product # MA5-42558) at 1:500 dilution for 45 minutes at room temperature. Goat anti-Mouse IgG - HRP Secondary Antibody was used at 1:100,000 dilution for 30 minutes at room temperature. Lane 1: Hela whole cell lysates at 4 μg; Lane 2: CENPB IP in Hela whole cell lysates; Lane 3: Mouse IgG instead of CENPB in Hela whole cell lysates. Predicted band size: 65 kDa; Observed band size: 85 kDa. Exposure time: 2 minutes; 8% SDS-PAGE gel.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry-Immunofluorescence analysis of of CENPB in A549 cells (green). Methanol fixed cells were blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with CENPB Monoclonal Antibody (Product # MA5-42558) at a dilution of 1:50 for 1 hour at room temperature, washed with PBS. Alexa Fluor 488 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1:1,000 dilution. The nuclear counter stain is DAPI (blue).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry (Paraffin) analysis of paraffin-embedded human kidney tissue using CENPB Monoclonal Antibody (Product # MA5-42558). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the CENPB antibody at a dilution of 1:400 for 30 minutes 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.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry (Paraffin) analysis of paraffin-embedded human skin tissue using CENPB Monoclonal Antibody (Product # MA5-42558). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the CENPB antibody at a dilution of 1:100 for 30 minutes 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.