Anti-GRO gamma antibody [EPR21759-11]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GRO gamma antibody [EPR21759-11] (AB220431)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, with 300 ng/ml Brefeldin A (BFA) added after 3 hours or untreated labeling GRO gamma with ab220431 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in RAW 264.7 cells treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, with 300 ng/ml Brefeldin A (BFA) added after 3 hours.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-GRO gamma antibody [EPR21759-11] (AB220431)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized RAW264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, with 300 ng/ml Brefeldin A (BFA) added after 3 hours (Right panel) or untreated (Left panel) labeling GRO gammawith ab220431 at 1/500 dilution. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

Note : CINC-2 and GRO gamma are alternative names for the same target.

• WB

Supplier Data

Western blot - Anti-GRO gamma antibody [EPR21759-11] (AB220431)

Exposure time : Lane 1 : 15 seconds; Lane 2 : 3 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

Observed MW : 9 kDa (lane 1), 11 kDa (lane 2).

The molecular mass observed is consistent with what has been described in the literature (PMID : 12508094).

Note : CINC-2 and GRO gamma are alternative names for the same target.

All lanes:

Western blot - Anti-GRO gamma antibody [EPR21759-11] (ab220431) at 1/1000 dilution

Lane 1:

Rat CINC-2 recombinant protein (aa33-100) at 0.01 µg

Lane 2:

Rat lung primary cell supernatant at 10 µL

Secondary

Lane 1:

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

Lane 2:

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 1/50000 dilution

Predicted band size: 11 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-GRO gamma antibody [EPR21759-11] (AB220431)

GRO gamma was immunoprecipitated from 0.35 mg of RAW264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, with 300 ng/ml Brefeldin A (BFA) added after 3 hours, whole cell lysate with ab220431 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220431 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : RAW264.7 treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, with 300 ng/ml Brefeldin A (BFA) added after 3 hours, whole cell lysate 10 μg (Input).

Lane 2 : ab220431 IP in RAW264.7 treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, with 300 ng/ml Brefeldin A (BFA) added after 3 hours, whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab220431 in RAW264.7 treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, with 300 ng/ml Brefeldin A (BFA) added after 3 hours, whole cell lysate.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-GRO gamma antibody [EPR21759-11] (ab220431)

Predicted band size: 11 kDa

false

• WB

Supplier Data

Western blot - Anti-GRO gamma antibody [EPR21759-11] (AB220431)

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression of CINC-2 increased LPS treatment of macrophages (PMID : 9576061).

Note : CINC-2 and GRO gamma are alternative names for the same target.

All lanes:

Western blot - Anti-GRO gamma antibody [EPR21759-11] (ab220431) at 1/1000 dilution

Lane 1:

Untreated RAW264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 2:

RAW264.7 treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, with 300ng/ml Brefeldin A (BFA) added after 3 hours whole cell lysate at 10 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 1/50000 dilution

Predicted band size: 11 kDa

Observed band size: 11 kDa

false

Exposure time: 3s