701070

antibody from Invitrogen Antibodies

Targeting: INSR CD220

Western blot (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

701070

Provider

Invitrogen Antibodies

Product name

Phospho-INSR (Tyr1158) Recombinant Rabbit Monoclonal Antibody (8H20L22)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

8H20L22

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Phospho-IR pTyr1158 in whole cell extracts of serum-starved HeLa cells treated with insulin (100 ng/mL, 15 min) using a Phospho-IR pTyr1158 recombinant rabbit monoclonal antibody (Product # 701070) at a dilution of 1 µg/mL. To confirm specificity, competition was performed by preincubation with the phosphopeptide to inhibit antibody binding (lane 2). Results show a band at ~115 kDa.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Phospho-IR pTyr1158 in whole cell extracts of serum-starved HeLa cells treated with insulin (100 ng/mL, 15 min) using a Phospho-IR pTyr1158 recombinant rabbit monoclonal antibody (Product # 701070) at a dilution of 1 µg/mL. To confirm specificity, competition was performed by preincubation with the phosphopeptide to inhibit antibody binding (lane 2). Results show a band at ~115 kDa.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of IR [pY1158] was done on MCF7 cells treated with Okadaic acid (100nM, 5 minutes) followed by Insulin (100nM, 10 minutes). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with ABfinity™ IR [pY1158] Recombinant Rabbit Monoclonal Antibody (701070, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.