Anti-XAF1 antibody [EPR26960-175] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-XAF1 antibody [EPR26960-175] - BSA and Azide free (AB313880)

This data was developed using ab313879, the same antibody clone in a different buffer formulation. XAF1 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) treated with 10 ng/ml human IFN alpha for 24 hours whole cell lysate with ab313879 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313879 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) treated with 10 ng/ml human IFN alpha for 24 hours whole cell lysate Lane 2 : ab313879 IP in HeLa (human cervical adenocarcinoma epithelial cell) treated with 10 ng/ml human IFN alpha for 24 hours whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313879 in HeLa treated with 10 ng/ml human IFN alpha for 24 hours whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 76 seconds

All lanes:

Immunoprecipitation - Anti-XAF1 antibody [EPR26960-175] (<a href='/en-us/products/primary-antibodies/xaf1-antibody-epr26960-175-ab313879'>ab313879</a>) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) treated with 10 ng/ml human IFN alpha for 24 hours whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 76s

• WB

Supplier Data

Western blot - Anti-XAF1 antibody [EPR26960-175] - BSA and Azide free (AB313880)

This data was developed using ab313879, the same antibody clone in a different buffer formulation.

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

Negative control : K-562 (PMID : 11175744).

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

Exposure time : Lanes 1-2 : 59 seconds; Lanes 3-5 : 6 seconds.

All lanes:

Western blot - Anti-XAF1 antibody [EPR26960-175] (<a href='/en-us/products/primary-antibodies/xaf1-antibody-epr26960-175-ab313879'>ab313879</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa treated with /ml human IFN alpha for 24 hours whole cell lysate at 20 µg

Lane 3:

Untreated Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 4:

Jurkat treated with /ml human IFN alpha overnight whole cell lysate at 20 µg

Lane 5:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 38 kDa,32 kDa

false