Anti-CXCL1/GRO alpha antibody [EPR23533-315] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-CXCL1/GRO alpha antibody [EPR23533-315] - BSA and Azide free (AB307590)

This data was developed using ab307589, the same antibody clone in a different buffer formulation. CXCL1/GRO alpha was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) treated with 0.5µg/ml Lipopolysaccharides(LPS) for 24 hours and then added 300 ng/ml Brefeldin A for another 20 hours, whole cell lysate 10 µg with ab307589 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab307589 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HUVEC treated with 0.5µg/ml Lipopolysaccharides(LPS) for 24 hours and then added 300 ng/ml Brefeldin A for another 20 hours, whole cell lysate 10 µg Lane 2 : ab307589 IP in HUVEC treated with 0.5µg/ml Lipopolysaccharides(LPS) for 24 hours and then added 300 ng/ml Brefeldin A for another 20 hours, whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307589 in HUVEC treated with 0.5µg/ml Lipopolysaccharides(LPS) for 24 hours and then added 300 ng/ml Brefeldin A for another 20 hours, whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-CXCL1/GRO alpha antibody [EPR23533-315] (<a href='/en-us/products/primary-antibodies/cxcl1-gro-alpha-antibody-epr23533-315-ab307589'>ab307589</a>) at 1/30 dilution

All lanes:

HUVEC treated with 0.5ug/ml Lipopolysaccharides

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

Observed band size: 11 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CXCL1/GRO alpha antibody [EPR23533-315] - BSA and Azide free (AB307590)

This data was developed using ab307589, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. This antibody does not cross-react with human CXCL2 and CXCL3. Exposure time : 180 seconds.

All lanes:

Western blot - Anti-CXCL1/GRO alpha antibody [EPR23533-315] (<a href='/en-us/products/primary-antibodies/cxcl1-gro-alpha-antibody-epr23533-315-ab307589'>ab307589</a>) at 1/1000 dilution

Lane 1:

His-tagged human CXCL1 recombinant protein at 10 ng

Lane 2:

His-tagged human CXCL2 recombinant protein at 10 ng

Lane 3:

His-tagged human CXCL3 recombinant protein at 10 ng

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

Observed band size: 11 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-CXCL1/GRO alpha antibody [EPR23533-315] - BSA and Azide free (AB307590)

This data was developed using ab307589, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-CXCL1/GRO alpha antibody [EPR23533-315] (<a href='/en-us/products/primary-antibodies/cxcl1-gro-alpha-antibody-epr23533-315-ab307589'>ab307589</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HUVEC treated with 0.5ug/ml Lipopolysaccharides (LPS) for 4 hours and then added 300 ng/ml Brefeldin A for another 20 hours 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>)

Predicted band size: 11 kDa

Observed band size: 11 kDa

false

Exposure time: 180s