Antibodies, Proteins, Kits and Reagents for Life Science

324 988 ₸ Product size

100 µl 324 988 ₸

Order now and get it on Tuesday March 02, 2021

Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19322] to IGF1 Receptor
Suitable for: WB, Flow Cyt, ICC/IF, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-IGF1 Receptor antibody [EPR19322]
See all IGF1 Receptor primary antibodies
Description

Rabbit monoclonal [EPR19322] to IGF1 Receptor
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse Rat
IP	Mouse
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: C2C12, HeLa, 293, MCF7, C6, and NIH/3T3 whole cell lysates; Mouse brain and spleen lysates; Rat brain lysate. ICC/IF: C2C12 and C6 cells. Flow Cyt: C2C12 cells. IP: C2C12 whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19322
Isotype

IgG
Research areas

Western blot - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

All lanes : Anti-IGF1 Receptor antibody [EPR19322] (ab182408) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : IGF1R knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 156 kDa
Observed band size: 100 kDa
why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab182408 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab182408 was shown to react with IGF1 Receptor in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264801 (knockout cell lysate ab256951) was used. Wild-type HeLa and IGF1R knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab182408 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

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 counter stain 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.
Western blot - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

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.
Western blot - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

Lanes 1-4 : Anti-IGF1 Receptor antibody [EPR19322] (ab182408) at 1/2000 dilution
Lanes 5-6 : Anti-IGF1 Receptor antibody [EPR19322] (ab182408) at 1/1000 dilution

Lane 1 : C2C12 (Mouse myoblast cell line) whole cell lysate at 20 µg
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 3 : 293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 5 : C6 (Rat glial tumor cell line) whole cell lysate at 10 µg
Lane 6 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 156 kDa
Observed band size: 100,200 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 5 seconds; Lane 2-4: 3 minutes; Lane 5/6: 2 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID: 11402025).
Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

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 counter stain 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.
Western blot - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

Lane 1 : Anti-IGF1 Receptor antibody [EPR19322] (ab182408) at 1/1000 dilution
Lanes 2-3 : Anti-IGF1 Receptor antibody [EPR19322] (ab182408) at 1/5000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse spleen lysate
Lane 3 : Rat brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 156 kDa
Observed band size: 100,200 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 10 seconds; Lane 2: 3 minutes; Lane 3: 1 minute.

The expression profile observed is consistent with what has been described in the literature (PMID: 11402025).
Flow Cytometry - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

Flow cytometric analysis of 4% paraformaldehyde-fixed 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 unlabelled 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.
Immunoprecipitation - Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

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.
Anti-IGF1 Receptor antibody [EPR19322] (ab182408)

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