Anti-GM-CSF antibody [EPR25991-223] (BSA and Azide free)

• IP

Supplier Data

Immunoprecipitation - Anti-GM-CSF antibody [EPR25991-223] (BSA and Azide free) (AB300496)

This data was developed using ab300495, the same antibody clone in a different buffer formulation.

CSF2 was immunoprecipitated from 0.35 mg HuT-78 (human Sezary syndrome cutaneous T lymphocyte) treated with 80nM TPA and 30μM ionomycin for 5 hours, then with 300ng/ml BFA added for 4 hours whole cell lysate 10 µg with ab300495 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300495 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HuT-78 (human Sezary syndrome cutaneous T lymphocyte) treated with 80nM TPA and 30μM ionomycin for 5 hours, then with 300ng/ml BFA added for 4 hours whole cell lysate 10 µg

Lane 2 : ab300495 IP in HuT-78 treated with 80nM TPA and 30μM ionomycin for 5 hours, then with 300ng/ml BFA added for 4 hours cell lysates

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300495 in HuT-78 treated with 80nM TPA and 30μM ionomycin for 5 hours, then with 300ng/ml BFA added for 4 hours cell lysates

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

Exposure time : 75 seconds

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 14512323).

All lanes:

Immunoprecipitation - Anti-GM-CSF antibody [EPR25991-223] (<a href='/en-us/products/primary-antibodies/gm-csf-antibody-epr25991-223-ab300495'>ab300495</a>) at 1/1000 dilution

All lanes:

HuT-78 treated with 80nM TPA and 30μM ionomycin for 5 hours, then with 300ng/ml BFA added for 4 hours cell lysates at 10 µg

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: 75s

• WB

Supplier Data

Western blot - Anti-GM-CSF antibody [EPR25991-223] (BSA and Azide free) (AB300496)

This data was developed using ab300495, the same antibody clone in a different buffer formulation. Blocking Buffer and concentration : 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 14512323).

All lanes:

Western blot - Anti-GM-CSF antibody [EPR25991-223] (<a href='/en-us/products/primary-antibodies/gm-csf-antibody-epr25991-223-ab300495'>ab300495</a>) at 1/1000 dilution

Lane 1:

Untreated HuT-78 (human Sezary syndrome cutaneous T lymphocyte) whole cell lysate

Lane 2:

HuT-78 treated with 80nM TPA and 30uM ionomycin for 5 hours, then with /ml BFA added for 4 hours whole cell lysate 20 µg

Lane 3:

Untreated Jurkat (human T cell leukemia T lymphocyte) whole cell lysate

Lane 4:

Jurkat treated with 80nM TPA and 30uM ionomycin for 5 hours, then with /ml BFA added for 4 hours whole cell lysate 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 1/100000 dilution

Observed band size: 15 kDa,18 kDa,22 kDa

false