Anti-EBI3 antibody [EPR28427-89] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-EBI3 antibody [EPR28427-89] - BSA and Azide free (AB316202)

This data was developed using ab316201, the same antibody clone in a different buffer formulation.

EBI3 was immunoprecipitated from 0.35 mg RAW 264.7 treated with 100 ng/ml LPS for 4 hours, 1 ug/ml BFA was then added for additional 3 hours, whole cell lysate with ab316201 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316201 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-EBI3 antibody [EPR28427-89] (<a href='/en-us/products/primary-antibodies/ebi3-antibody-epr28427-89-ab316201'>ab316201</a>) at 1/1000 dilution

Lane 1:

RAW 264.7 treated with 100 ng/ml LPS for 4 hours, 1 ug/ml BFA was then added for additional 3 hours, whole cell lysate

Lane 2:

<a href='/en-us/products/primary-antibodies/ebi3-antibody-epr28427-89-ab316201'>ab316201</a> at 1/30 IP in RAW 264.7 treated with 100 ng/ml LPS for 4 hours, 1 ug/ml BFA was then added for additional 3 hours, whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/ebi3-antibody-epr28427-89-ab316201'>ab316201</a> in RAW 264.7 treated with 100 ng/ml LPS for 4 hours, 1 ug/ml BFA was then added for additional 3 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: 180s

• WB

Supplier Data

Western blot - Anti-EBI3 antibody [EPR28427-89] - BSA and Azide free (AB316202)

This data was developed using ab316201, the same antibody clone in a different buffer formulation.

The up-regulation of EBI3 is induced by LPS treatment (PMID : 15728491).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-EBI3 antibody [EPR28427-89] (<a href='/en-us/products/primary-antibodies/ebi3-antibody-epr28427-89-ab316201'>ab316201</a>) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 treated with /ml LPS for 4 hours, 1/ml BFA was then added for additional 3 hours, whole cell lysate µg 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: 30 kDa,36 kDa

false

Exposure time: 180s