Anti-IGF1 Receptor antibody [EPR19322] - Low endotoxin, Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR19322] - Low endotoxin, Azide free (AB246702)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (Mouse myoblast cell line) cells labeling IGF1 Receptor with ab182408 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on C2C12 cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows :

-ve control 1 : ab182408 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 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 (ab182408).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR19322] - Low endotoxin, Azide free (AB246702)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cell line) cells labeling IGF1 Receptor with ab182408 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on C6 cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows :

-ve control 1 : ab182408 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 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 (ab182408).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IGF1 Receptor antibody [EPR19322] - Low endotoxin, Azide free (AB246702)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized C2C12 (Mouse myoblast cell line) cells labeling IGF1 Receptor with ab182408 at 1/80 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 (Alexa Fluor^® 488) at 1/500 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 (ab182408).

• IP

Supplier Data

Immunoprecipitation - Anti-IGF1 Receptor antibody [EPR19322] - Low endotoxin, Azide free (AB246702)

IGF1 Receptor was immunoprecipitated from 1mg of C2C12 (Mouse myoblast cell line) whole cell lysate with ab182408 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab182408 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : C2C12 whole cell lysate, 10μg (Input).

Lane 2 : ab182408 IP in C2C12 whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab182408 in C2C12 whole cell lysate.

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

Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab182408).

All lanes:

Immunoprecipitation - Anti-IGF1 Receptor antibody [EPR19322] (<a href='/en-us/products/primary-antibodies/igf1-receptor-antibody-epr19322-ab182408'>ab182408</a>)

Predicted band size: 154 kDa

false

• WB

Lab

Western blot - Anti-IGF1 Receptor antibody [EPR19322] - Low endotoxin, Azide free (AB246702)

Lane 1 : Wild-type HAP1 whole cell lysate (40 μg)
Lane 2 : IGF1R knockout HAP1 whole cell lysate (40 μg)
Lane 3 : HeLa whole cell lysate (40 μg)

Lanes 1 - 3 Merged signal (red and green). Green - ab182408 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab182408 was shown to specifically recognize IGF1R in wild-type HAP1 cells along with additional cross-reactve bands. No band was observed when IGF1R knockout samples were examined. Wild-type and IGF1R knockout samples were subjected to SDS-PAGE. ab182408 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab182408).

All lanes:

Western blot - Anti-IGF1 Receptor antibody [EPR19322] (<a href='/en-us/products/primary-antibodies/igf1-receptor-antibody-epr19322-ab182408'>ab182408</a>)

Predicted band size: 154 kDa

false

• WB

Lab

Western blot - Anti-IGF1 Receptor antibody [EPR19322] - Low endotoxin, Azide free (AB246702)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab182408). Anti-IGF1R antibody [EPR19322] (ab182408) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab182408 was shown to bind specifically to IGF1R. A band was observed at 82 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in IGF1R knockout cell line. To generate this image, wild-type and IGF1R knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-IGF1 Receptor antibody [EPR19322] (<a href='/en-us/products/primary-antibodies/igf1-receptor-antibody-epr19322-ab182408'>ab182408</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot at 20 µg

Lane 2:

Western blot - Human IGF1R knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-igf1r-knockout-hct116-cell-line-ab287509'>ab287509</a>)

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 154 kDa

Observed band size: 82 kDa

false