Anti-p115-RhoGEF antibody [JH-1] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-p115-RhoGEF antibody [JH-1] - BSA and Azide free (AB255755)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized EL4 (mouse lymphoma T lymphocyte) cells labelling p115-RhoGEF with ab243248 at 1/50 dilution, followed by ab173003 Goat Anti-Armenian hamster IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in EL4 cells. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control 1 : ab243248 at a 1/50 dilution followed by ab150080 at a 1/1000 dilution.

Negative control 2 : ab179513 at a 1/200 dilution followed by ab173003 at a 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243248).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-p115-RhoGEF antibody [JH-1] - BSA and Azide free (AB255755)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A20 (mouse reticulum sarcoma B lymphocyte) cells labelling p115-RhoGEF with ab243248 at 1/50 dilution, followed by ab173003 Goat Anti-Armenian hamster IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in A20 cells. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control 1 : ab243248 at a 1/50 dilution followed by ab150080 at a 1/1000 dilution.

Negative control 2 : ab179513 at a 1/200 dilution followed by ab173003 at a 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243248).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-p115-RhoGEF antibody [JH-1] - BSA and Azide free (AB255755)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized EL-4 (mouse lymphoma T lymphocyte) cells labelling p115-RhoGEF with ab243248 at 1/40 dilution (1μg) (Red) compared with an Armenian hamster monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti armenian hamster IgG (Alexa Fluor^® 488, ab173003) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide ab243248.

• WB

Lab

Western blot - Anti-p115-RhoGEF antibody [JH-1] - BSA and Azide free (AB255755)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11384980).

Lysates should be made freshly and used in WB immediately.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1/2：3 seconds; Lane 3：15 seconds; Lane 4：37 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243248).

All lanes:

Western blot - Anti-p115-RhoGEF antibody [JH-1] (<a href='/en-us/products/primary-antibodies/p115-rhogef-antibody-jh-1-ab243248'>ab243248</a>) at 1/1000 dilution

Lane 1:

A20 (mouse reticulum sarcoma B lymphocyte), whole cell lysate at 10 µg

Lane 2:

EL4 (mouse lymphoma T lymphocyte), whole cell lysate at 10 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Lane 4:

C6 (rat glial tumor glial cell), whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Rabbit Anti-Armenian hamster IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/rabbit-armenian-hamster-igg-h-l-hrp-ab5745'>ab5745</a>) at 1/5000 dilution

Predicted band size: 102 kDa

Observed band size: 115 kDa

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