701053

antibody from Invitrogen Antibodies

Targeting: EGFR ERBB, ERBB1

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

701053

Provider

Invitrogen Antibodies

Product name

Anti-Phospho-EGFR (Tyr1068) Antibody (2H9L3), ABfinity™ Rabbit Monoclonal

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with human based on sequence homology. ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A. ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance. Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

2H9L3

Vial size

100 µg

Concentration

0.5 mg/ml

Storage

Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C

References

Submitted references

Evaluation of dimerization-inhibitory activities of cyclic peptides containing a β-hairpin loop sequence of the EGF receptor.

Mizuguchi T, Ohara N, Iida M, Ninomiya R, Wada S, Kiso Y, Saito K, Akaji K
Bioorganic & medicinal chemistry 2012 Oct 1;20(19):5730-7

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of EGFR (pY1068) was performed on 70% confluent log phase A549 cells. 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 ABfinity™ EGFR (pY1068) recombinant rabbit monoclonal antibody (Product # 701053) at a dilution of 1:500 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 cytoplasmic localization of phosphorylated EGFR (pY1068). Panel e shows competition with the phospho-AKT (pT308) peptide. The images were captured using a Nikon microscope at 20X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of EGFR (pY1068) was performed on A549 cells treated with EGF (100 ng/ml, 5 minutes). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0. 25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were labeled with ABfinityª EGFR (pY1068) recombinant rabbit monoclonal antibody (Product # 701053, red histogram) or with rabbit isotype control (pink histogram) at a dilution of 1:250 in 2.5% BSA. After incubation at room temperature for 3 hours, the cells were labeled with Alexa Fluor¨ 488 goat anti-Rabbit Secondary antibody (Product # 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.