Revision 1

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-EGF Receptor (Tyr1068) (D7A5) Rabbit Monoclonal Antibody3777100 µl175 kDaRabbit IgG
EGF Receptor (D38B1) Rabbit Monoclonal Antibody4267100 µl175 kDaRabbit IgG

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

UniProt ID:
#P00533

Entrez-Gene Id:
1956

Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, and lysosomal degradation (1,2). Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme, and providing a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of EGFR at Tyr845 (5). The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling (6). Phosphorylation of EGFR at Tyr1045 creates a major docking site for the adaptor protein c-Cbl, leading to receptor ubiquitination and degradation following EGFR activation (7,8). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (9). A pair of phosphorylated EGFR residues (Tyr1148 and Tyr1173) provide a docking site for the Shc scaffold protein, with both sites involved in MAP kinase signaling activation (2). Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PhosphoPlus is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from serum-starved A431 cells, untreated (-) or treated with hEGF (100 ng/ml, 5 min; +), using EGF Receptor (1F4) Mouse mAb #2239 (Panel A) and Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777 (Panel B). Anti-rabbit IgG (H+L) (DyLight 800 4X PEG Conjugate) #5151 (green) and Anti-mouse IgG (H+L) (DyLight 680 Conjugate) #5470 (red) were used as secondary antibodies.
undefined Image 1: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Western blot analysis of extracts from control Hela cells (lane 1), or EGFR knockout Hela cells (lane 2) using EGF Receptor (D38B1) XP® Rabbit mAb #4267, (upper) or #8457 β-Actin (D6A8) Rabbit mAb (lower). The absence of signal in EGFR-knockout Hela cells confirms specificity of the antibody for EGFR.
undefined Image 2: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using EGF Receptor (D38B1) Rabbit mAb performed on the Leica® BOND™ Rx.
undefined Image 3: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts of BxPC-3 cells, untreated or EGF-stimulated, using Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb (upper) and EGF Receptor Antibody #2232 (lower).
undefined Image 4: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Immunohistochemical analysis of paraffin-embedded HCC827 xenograft, control (left) or λ phosphatase-treated (right), using Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb.
undefined Image 5: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Western blot analysis of extracts from A-431, BxPC3 and HeLa cells using EGF Receptor (D38B1) XP® Rabbit mAb.
undefined Image 6: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using EGF Receptor (D38B1) XP® Rabbit mAb.
undefined Image 7: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Immunohistochemical analysis using Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb on SignalSlide™ Phospho-EGF Receptor IHC Controls #8102 (paraffin-embedded KYSE450 cell pellets, untreated (left) or EGF-treated (right)).
undefined Image 8: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Simple Western™ analysis of lysates (0.1 mg/mL) from A431 cells treated with human EGF (100 ng/mL, 30”) using Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777. The virtual lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 9: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using EGF Receptor (D38B1) XP® Rabbit mAb.
undefined Image 10: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human placenta using EGF Receptor (D38B1) XP® Rabbit mAb.
undefined Image 11: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or EGF-treated (right), using Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb (green) and DRAQ5® #4084 (blue pseudocolor).
undefined Image 12: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded rhesus monkey kidney using EGF Receptor (D38B1) Rabbit mAb.
undefined Image 13: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Flow cytometric analysis of A549 cells, untreated (blue) or EGF-treated (green), using Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb compared to concentration matched XP® Rabbit (DA1E) mAb IgG Isotype Control #3900 (red).
undefined Image 14: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Immunohistochemical analysis of paraffin-embedded MDA-MB-468 (amplified EGFR, left), HT-29 (low EGFR, middle) and CAMA-1 (EGFR negative, right) cells using EGF Receptor (D38B1) XP® Rabbit mAb.
undefined Image 15: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of A549 cells, untreated (left) or treated with human epidermal growth factor (right), using EGF Receptor (D38B1) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 16: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Flow cytometric analysis of Jurkat cells (blue) and A431 cells (green) using EGF Receptor (D38B1) XP® Rabbit mAb #4267 (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 17: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Simple Western™ analysis of lysates (0.1 mg/mL) from A-431 cells using EGF Receptor (D38B1) XP® Rabbit mAb #4267. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
undefined Image 18: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunoprecipitation of EGFR protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is EGF Receptor (D38B1) XP® Rabbit mAb. Western blot analysis was performed using EGF Receptor (D38B1) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
undefined Image 19: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Western blot analysis of extracts from HeLa, mIMCD-3, and Cos-7 cells using EGF Receptor (D38B1) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
undefined Image 20: PhosphoPlus<sup>®</sup> EGFR (Tyr1068) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.