AF1095

antibody from R&D Systems

Targeting: EGFR ERBB, ERBB1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

AF1095

Provider

R&D Systems

Product name

Human Phospho-EGFR (Y1173) Antibody

Antibody type

Polyclonal

Description

Antigen Affinity-purified. Detects human EGFR when phosphorylated at Y1173 in Western blots.

Reactivity

Human

Host

Rabbit

Conjugate

Unconjugated

Isotype

IgG

Vial size

50 ug

Concentration

LYOPH

Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution.

References

Submitted references

ADAM17 substrate release in proximal tubule drives kidney fibrosis.

Kefaloyianni E, Muthu ML, Kaeppler J, Sun X, Sabbisetti V, Chalaris A, Rose-John S, Wong E, Sagi I, Waikar SS, Rennke H, Humphreys BD, Bonventre JV, Herrlich A
JCI insight 2016 Aug 18;1(13)

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Phenotypic diversity of breast cancer-related mutations in metalloproteinase-disintegrin ADAM12.

Qi Y, Duhachek-Muggy S, Li H, Zolkiewska A
PloS one 2014;9(3):e92536

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Analysis of Epithelial Growth Factor-Receptor (EGFR) Phosphorylation in Uterine Smooth Muscle Tumors: Correlation to Mucin-1 and Galectin-3 Expression.

Weissenbacher T, Vrekoussis T, Roeder D, Makrigiannakis A, Mayr D, Ditsch N, Friese K, Jeschke U, Dian D
International journal of molecular sciences 2013 Feb 28;14(3):4783-92

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Protein C is an autocrine growth factor for human skin keratinocytes.

Xue M, Campbell D, Jackson CJ
The Journal of biological chemistry 2007 May 4;282(18):13610-6

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Repulsion of cerebellar granule neurons by chondroitin sulfate proteoglycans is mediated by MAPK pathway.

Kaneko M, Kubo T, Hata K, Yamaguchi A, Yamashita T
Neuroscience letters 2007 Aug 9;423(1):62-7

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human Phospho-EGFR (Y1173) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 μM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 0.2 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody, followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-EGFR (Y1173) at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human Phospho-EGFR (Y1173) by Simple Western^TM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-EGFR (Y1173) at approximately 265 kDa (as indicated) using 2 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.

Immunocytochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Phospho-EGFR (Y1173) in A431 Human Cell Line. EGFR phosphorylated at Y1173 was detected in immersion fixed A431 human epithelial carcinoma cell line untreated (lower panel) or treated (upper panel) with pervanadate using Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Phospho-EGFR (Y1173) in Mouse Embryo. EGFR phosphorylated at Y1173 was detected in immersion fixed frozen sections of mouse embryo using Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Flow cytometry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Phospho-EGFR (Y1173) in A431 Human Cell Line by Flow Cytometry. A431 human epithelial carcinoma cells were untreated (open histogram), or treated for 5 minutes with 100 ng/mL Recombinant Human EGF (Catalog # 236-EG; filled histogram) then stained with Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with para-formaldehyde and permeabilized with saponin.