Anti-BAFF antibody [EPR19704] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BAFF antibody [EPR19704] - BSA and Azide free (AB251400)

This data was developed using ab203791, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T (Human epithelial cell line from embryonic kidney) cells labeling BAFF with ab203791 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 647) (ab150079) secondary antibody at 1/1000 dilution.

Confocal image showing positive staining on 293T cells transfected with a GFP-tagged BAFF expression construct.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150079 at 1/1000 dilution.

ICC/IF was also performed on a few cell lines including U937, no signal was detected.

• IP

Supplier Data

Immunoprecipitation - Anti-BAFF antibody [EPR19704] - BSA and Azide free (AB251400)

This data was developed using ab203791, the same antibody clone in a different buffer formulation.

BAFF was immunoprecipitated from 0.35 mg of U937 (Human histiocytic lymphoma cell line) whole cell lysate with ab203791 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab203791 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : U937 whole cell lysate, 10μg (Input).
Lane 2 : ab203791 IP in U937 whole cell lysate.
Lane 3 : Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab203791 in U937 whole cell lysate.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-BAFF antibody [EPR19704] (<a href='/en-us/products/primary-antibodies/baff-antibody-epr19704-ab203791'>ab203791</a>)

Predicted band size: 31 kDa

false

• WB

Supplier Data

Western blot - Anti-BAFF antibody [EPR19704] - BSA and Azide free (AB251400)

This data was developed using ab203791, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Two isoforms have been described in literature (PMID : 12867412).

All lanes:

Western blot - Anti-BAFF antibody [EPR19704] (<a href='/en-us/products/primary-antibodies/baff-antibody-epr19704-ab203791'>ab203791</a>) at 1/1000 dilution

All lanes:

U937 (Human histiocytic lymphoma cell line) whole cell lysate at 10 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

Predicted band size: 31 kDa

Observed band size: 17 kDa,29 kDa,31 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-BAFF antibody [EPR19704] - BSA and Azide free (AB251400)

This data was developed using ab203791, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-BAFF antibody [EPR19704] (<a href='/en-us/products/primary-antibodies/baff-antibody-epr19704-ab203791'>ab203791</a>) at 1/5000 dilution

Lane 1:

Empty vector with GFP-Myc tag (vector control) transfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg

Lane 2:

HEK-293 (Human epithelial cell line from embryonic kidney) transfected with a GFP-Myc tagged BAFF vector whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 31 kDa

Observed band size: 57 kDa

false

Exposure time: 1s