PA5-80234

antibody from Invitrogen Antibodies

Targeting: XAF1 BIRC4BP, HSXIAPAF1, XIAPAF1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-80234

Provider

Invitrogen Antibodies

Product name

XAF1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL. Positive Control - WB: human Jurkat whole cell. ICC/IF: U2OS cell. Flow: SiHa cell.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

500 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of XAF1 in Lane 1: HEPG2 whole cell lysate, Lane 2: HEPG2 whole cell lysate using 40-50 µg per well. Sample was incubated with XAF1 (Product # PA5-80234) at a dilution of 0.5 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of XAF1 using XAF1 Polyclonal Antibody (Product # PA5-80234). Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel)/120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. Lane 1: human Jurkat whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with XAF1 Polyclonal Antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate with Tanon 5200 system. A specific band was detected for XAF1 at approximately 35 kDa. The expected band size for XAF1 is at 35 kDa.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry/Immunofluorescence analysis of XAF1 in U2OS cells using XAF1 Polyclonal Antibody (Product # PA5-80234). Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and incubated with the primary antibody at 5 µg/mL. DyLight 488 conjugated goat anti-rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualized using a fluorescence microscope and filter sets appropriate for the label used.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of XAF1 on paraffin-embedded human Endometrial Carcinoma tissue. Sample was incubated with XAF1 polyclonal antibody (Product# PA5-80234).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of XAF1 on paraffin-embedded human Endometrial Carcinoma tissue. Sample was incubated with XAF1 polyclonal antibody (Product# PA5-80234).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of XAF1 in SiHa cells using XAF1 Polyclonal Antibody (Product # PA5-80234), shown in overlay histogram (blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum, and incubated with the primary antibody (1 μg/1x10^6 cells) for 30 min at 20°C. DyLight 488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.