Anti-IRS1+IRS2 antibody [EP263Y] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS1+IRS2 antibody [EP263Y] - BSA and Azide free (AB247286)

This data was developed using ab40777, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma labelling IRS1 with unpurified ab40777.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS1+IRS2 antibody [EP263Y] - BSA and Azide free (AB247286)

This data was developed using ab40777, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling IRS1 with purified ab40777 at a dilution of 1/100. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-IRS1+IRS2 antibody [EP263Y] - BSA and Azide free (AB247286)

This data was developed using ab40777, the same antibody clone in a different buffer formulation.

Intracellular Flow Cytometry analysis of MCF-7 (human breast carcinoma) cells labeling IRS1 with unpurified ab40777 at 1/90 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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-IRS1+IRS2 antibody [EP263Y] - BSA and Azide free (AB247286)

This data was developed using ab40777, the same antibody clone in a different buffer formulation.

Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling IRS1 with purified ab40777 at a dilution of 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1 : primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2 : ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

• WB

Unknown

Western blot - Anti-IRS1+IRS2 antibody [EP263Y] - BSA and Azide free (AB247286)

This data was developed using ab40777, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-IRS1+IRS2 antibody [EP263Y] (<a href='/en-us/products/primary-antibodies/irs1irs2-antibody-ep263y-ab40777'>ab40777</a>) at 1/1000 dilution

All lanes:

HeLa cell lysate at 10 µg

Observed band size: 170 kDa

false

• WB

Lab

Western blot - Anti-IRS1+IRS2 antibody [EP263Y] - BSA and Azide free (AB247286)

This data was developed using ab40777, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-IRS1+IRS2 antibody [EP263Y] (<a href='/en-us/products/primary-antibodies/irs1irs2-antibody-ep263y-ab40777'>ab40777</a>) at 1/10000 dilution

All lanes:

HEK293 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 1/20000 dilution

Observed band size: 170 kDa

false

• WB

Lab

Western blot - Anti-IRS1+IRS2 antibody [EP263Y] - BSA and Azide free (AB247286)

This data was developed using ab40777, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-IRS1+IRS2 antibody [EP263Y] (<a href='/en-us/products/primary-antibodies/irs1irs2-antibody-ep263y-ab40777'>ab40777</a>)

All lanes:

HeLa 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 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 170 kDa

false

• WB

Supplier Data

Western blot - Anti-IRS1+IRS2 antibody [EP263Y] - BSA and Azide free (AB247286)

This data was developed using ab40777, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM /TBST. Exposure time : Lane 1, 2, 4, 5 , 6 : 1 second. Lane 3 : 80 seconds.

Lanes 1 - 3:

Western blot - Anti-IRS1+IRS2 antibody [EP263Y] (<a href='/en-us/products/primary-antibodies/irs1irs2-antibody-ep263y-ab40777'>ab40777</a>) at 1/1000 dilution

Lanes 4 - 6:

Western blot - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [EPR20018-251] (<a href='/en-us/products/primary-antibodies/ddddk-tag-binds-to-flag-tag-sequence-antibody-epr20018-251-ab205606'>ab205606</a>) at 1/1000 dilution

Lanes 1 and 4:

HEK-293T transfected with an empty vector whole cell lysate at 20 µg

Lanes 2 and 5:

HEK-293T transfected with a human IRS1 expression vector containing a Myc/DDK tag whole cell lysate at 20 µg

Lanes 3 and 6:

HEK-293T transfected with a human IRS2 expression vector containing a Myc/DDK tag 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 1/20000 dilution

Observed band size: 170 kDa

false