Anti-ICOS Ligand/ICOSL antibody [EPR20240] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-ICOS Ligand/ICOSL antibody [EPR20240] - BSA and Azide free (AB251502)

This data was developed using ab209262, the same antibody clone in a different buffer formulation.

ICOS Ligand/ICOSL was immunoprecipitated from 0.35 mg of JAR (Human placenta choriocarcinoma cell line) whole cell lysate with ab209262 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab209262 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : JAR whole cell lysate, 10 μg (Input).

Lane 2 : ab209262 IP in JAR whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab209262 in JAR whole cell lysate.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-ICOS Ligand/ICOSL antibody [EPR20240] (<a href='/en-us/products/primary-antibodies/icos-ligand-icosl-antibody-epr20240-ab209262'>ab209262</a>)

Predicted band size: 33 kDa

Observed band size: 60 kDa

false

• WB

Supplier Data

Western blot - Anti-ICOS Ligand/ICOSL antibody [EPR20240] - BSA and Azide free (AB251502)

This data was developed using ab209262, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

The level of ICOS Ligand/ICOSL expression can be elevated by TNF-alpha treatment (PMID : 11983910).

All lanes:

Western blot - Anti-ICOS Ligand/ICOSL antibody [EPR20240] (<a href='/en-us/products/primary-antibodies/icos-ligand-icosl-antibody-epr20240-ab209262'>ab209262</a>) at 1/1000 dilution

Lane 1:

Untreated HUVEC (Human umbilical vein endothelial cell line) whole cell lysate at 20 µg

Lane 2:

HUVEC whole cell lysate treated with tumor necrosis factor-α (TNF-alpha, <a href='/en-us/products/proteins-peptides/recombinant-human-tnf-alpha-protein-ab9642'>ab9642</a>) 200 unit/ml for 24 hours 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

Predicted band size: 33 kDa

Observed band size: 60 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-ICOS Ligand/ICOSL antibody [EPR20240] - BSA and Azide free (AB251502)

This data was developed using ab209262, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

ICOS ligand/ICOSL is highly glycosylated (PMID 11983910).

Observed band sizes : 60 kDa (glycosylated form); 35 kDa (non-glycosylated form).

All lanes:

Western blot - Anti-ICOS Ligand/ICOSL antibody [EPR20240] (<a href='/en-us/products/primary-antibodies/icos-ligand-icosl-antibody-epr20240-ab209262'>ab209262</a>) at 1/1000 dilution

Lane 1:

Untreated JAR (Human placenta choriocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

JAR whole cell lysate treated with PNGase F 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

Predicted band size: 33 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-ICOS Ligand/ICOSL antibody [EPR20240] - BSA and Azide free (AB251502)

This data was developed using ab209262, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ICOS Ligand/ICOSL antibody [EPR20240] (<a href='/en-us/products/primary-antibodies/icos-ligand-icosl-antibody-epr20240-ab209262'>ab209262</a>) at 1/20000 dilution

All lanes:

His-tagged recombinant human ICOS ligand/ICOSL protein at 0.01 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: 33 kDa

Observed band size: 50 kDa

false

Exposure time: 10s