Anti-IRG1 antibody [EPR22066] - BSA and Azide free

• WB

Unknown

Western blot - Anti-IRG1 antibody [EPR22066] - BSA and Azide free (AB238169)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.

Lanes 1 & 2 : 3 minutes.

Lanes 3 & 4 : 8 seconds.

The molecular mass observed is consistent with what has been described in the literature (PMID : 23610393 and PMID : 28699638).
The protein is up-regulated after lipopolysaccharide (LPS) stimulation (PMID : 25640654 and PMID : 26679997).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab222411).

All lanes:

Western blot - Anti-IRG1 antibody [EPR22066] (<a href='/en-us/products/primary-antibodies/irg1-antibody-epr22066-ab222411'>ab222411</a>) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 (treated with 100 ng/ml lipopolysaccharide (LPS) for 4 hours) whole cell lysate at 20 µg

Lane 3:

Untreated THP-1 (human monocytic leukemia cell line) whole cell lysate at 20 µg

Lane 4:

THP-1 treated with 100 ng/ml phorbol-12-myristate-13-acetate (PMA) for 72 hours, and 1 μg/ml LPS for 16 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>) at 1/100000 dilution

Predicted band size: 53 kDa

Observed band size: 50 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-IRG1 antibody [EPR22066] - BSA and Azide free (AB238169)

IRG1 was immunoprecipitated from 0.35 mg of THP-1 (human monocytic leukemia cell line)(treated with 100 ng/ml phorbol-12-myristate-13-acetate (PMA) for 72 hours, then with 1 μg/ml LPS added for 16 hours) whole cell lysate with ab222411 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab222411 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

Lane 1 : THP-1 (treated with PMA/LPS) whole cell lysate, 10 μg (Input).
Lane 2 : ab222411 IP in THP-1 (treated with PMA/LPS) whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab222411 in THP-1 (treated with PMA/LPS) whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 8 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab222411).

All lanes:

Immunoprecipitation - Anti-IRG1 antibody [EPR22066] (<a href='/en-us/products/primary-antibodies/irg1-antibody-epr22066-ab222411'>ab222411</a>)

Predicted band size: 53 kDa

Observed band size: 50 kDa

false