MA5-18047

antibody from Invitrogen Antibodies

Targeting: CYBA p22-PHOX

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-18047

Provider

Invitrogen Antibodies

Product name

CYBA Monoclonal Antibody (CS9)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

This antibody is specific for human Cytochrome b245 light chain (aa 165-169, KKPSE).

Antibody clone number

CS9

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of CYBA/Cytochrome b245 Light Chain in human spleen cells using CYBA/Cytochrome b245 Light Chain monoclonal antibody (Product # MA5-18047).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of CYBA was performed using 70% confluent log phase PC-3, DU 145 and BJ cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CYBA Monoclonal Antibody (CS9) (Product # MA5-18047) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2500 dilution), for 45 minutes at room temperature (Panel a,e,i: Green). Nuclei (Panel b,f,j: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c,g,k: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d, h and i represents the merged image showing punctate staining in nucleus in both PC-3 and DU 145 but not in BJ. The images were captured at 60X magnification. CYBA localizes with Nox proteins in the nucleus in response to ROS. Between the prostate cancer cell lines, PC-3 constitutively produces higher ROS compared to DU 145 while BJ, a normal fibroblast cell line, has low endogenous levels [doi: 10.1158/0008-5472.CAN-07-5259; 10.1161/01.ATV.0000112024.13727.2c; 10.1089/ars.2008.2203]