Anti-IQGAP1 antibody [EPR5221] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-IQGAP1 antibody [EPR5221] - BSA and Azide free (AB247821)

This data was developed using ab109292, the same antibody clone in a different buffer formulation.

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling IQGAP1 with unpurified ab109292 at 1/250 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti-rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• WB

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Western blot - Anti-IQGAP1 antibody [EPR5221] - BSA and Azide free (AB247821)

This data was developed using ab109292, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-IQGAP1 antibody [EPR5221] - BSA and Azide free (ab247821) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

Cos-1 cell lysate at 10 µg

Lane 3:

HUVEC cell lysate at 10 µg

Lane 4:

Jurkat cell lysate at 10 µg

Predicted band size: 189 kDa

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• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-IQGAP1 antibody [EPR5221] - BSA and Azide free (AB247821)

This data was developed using ab109292, the same antibody clone in a different buffer formulation.

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling IQGAP1 with Purified ab109292 at 1/500 dilution (5 μg/ml). Cells were fixed with 100% methanol. ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

• WB

Lab

Western blot - Anti-IQGAP1 antibody [EPR5221] - BSA and Azide free (AB247821)

This data was developed using ab109292, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 cell lysate (20 μg)

Lane 2 : IQGAP1 knockout HAP1 cell lysate (20 μg)

Lane 3 : HEK293 cell lysate (20 μg)

Lane 4 : HeLa cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab109292 observed at 190 kDa. Red - loading control, ab7291, observed at 52 kDa.

ab109292 was shown to specifically react with IQGAP1 when IQGAP1 knockout samples were used. Wild-type and IQGAP1 knockout samples were subjected to SDS-PAGE. ab109292 and ab7291 (loading control to alpha tubulin) were diluted 1/1000 and 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-IQGAP1 antibody [EPR5221] (<a href='/en-us/products/primary-antibodies/iqgap1-antibody-epr5221-ab109292'>ab109292</a>)

Predicted band size: 189 kDa

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