Anti-GPNMB antibody [EPR18226-147] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GPNMB antibody [EPR18226-147] - BSA and Azide free (AB234529)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized L-929 (mouse connective tissue fibroblast cell line) and EL4 (mouse lymphoma T lymphocyte cell line) cells labeling GPNMB with ab188222 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 L-929 cells.

Negative control : EL4 (PMID : 19320736).

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188222).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-GPNMB antibody [EPR18226-147] - BSA and Azide free (AB234529)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized EL4 (mouse lymphoma T lymphocyte cell line; left panel) and L-929 (mouse connective tissue fibroblast cell line; right panel) cell lines labeling GPNMB with ab188222 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : EL4 (PMID : 19320736) (left panel).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188222).

• IP

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Immunoprecipitation - Anti-GPNMB antibody [EPR18226-147] - BSA and Azide free (AB234529)

GPNMB was immunoprecipitated from 0.35 mg of L-929 (mouse connective tissue fibroblast cell line) whole cell lysate with ab188222 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab188222 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : L-929 whole cell lysate 10 μg (Input).

Lane 2 : ab188222 IP in L-929 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab188222 in L-929 whole cell lysate.

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

Exposure time : 1 minute.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188222).

All lanes:

Immunoprecipitation - Anti-GPNMB antibody [EPR18226-147] (<a href='/en-us/products/primary-antibodies/gpnmb-antibody-epr18226-147-ab188222'>ab188222</a>)

Predicted band size: 64 kDa

Observed band size: 65 kDa,95 kDa

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