44-802G

antibody from Invitrogen Antibodies

Targeting: INSR CD220

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Antibody Data

Product number

44-802G

Provider

Invitrogen Antibodies

Product name

Phospho-IR/IGF1R (Tyr1158) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated insulin/insulin-like growth factor-1 receptor (IR/IGF1R). The final product is generated by affinity chromatography using an IR/IGF1R-derived peptide that is phosphorylated at tyrosine 1158 (tyrosine 1131 for IGF1R). CHO-T transfected with a vector containing insulin receptor and stimulated with insulin was used as positive control. The immunogenic sequence is conserved in mouse and rat for both the IR and IGF1R; this antibody is expected to react in human, mouse, and rat, and has been used in western blotting and immunostaining.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

IR/IGF1R (pY1158) and IR/IGF1R (pYpY1162/63) phosphospecific antibodies. CHO-T cell extracts transfected with an IR vector either unstimulated (-) or stimulated (+) with insulin (Product # 55150A) were immunoblotted with phosphospecific antibodies to IR/IGF1R. IR/IGF1R (pY1158) (1, 3). IR/IGF1R (pYpY1162/1163) (2, 4).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

IR/IGF1R (pY1158) and IR/IGF1R (pYpY1162/63) phosphospecific antibodies. CHO-T cell extracts transfected with an IR vector either unstimulated (-) or stimulated (+) with insulin (Product # 55150A) were immunoblotted with phosphospecific antibodies to IR/IGF1R. IR/IGF1R (pY1158) (1, 3). IR/IGF1R (pYpY1162/1163) (2, 4).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

IR/IGF1R (pY1158) and IR/IGF1R (pYpY1162/63) phosphospecific antibodies. CHO-T cell extracts transfected with an IR vector either unstimulated (-) or stimulated (+) with insulin (Product # 55150A) were immunoblotted with phosphospecific antibodies to IR/IGF1R. IR/IGF1R (pY1158) (1, 3). IR/IGF1R (pYpY1162/1163) (2, 4).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of IR/IGF1R (pY1158) was done on 70% confluent log phase MCF7 cells treated with insulin (100nM for 5 min). The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with IR/IGF1R (pY1158) Rabbit polyclonal Antibody (Product # 44-802G) at 2 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing membrane localization. Panel e shows untreated MCF7 cells. Panel f shows no primary antibody control. The images were captured at 20X magnification.