Revision 6
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, W-S, IP, IHC-P

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

185

Source/Isotype:

Rabbit 

UniProt ID:

#P21860

Entrez-Gene Id:

2065

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:1000

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.

For a carrier free (BSA and azide free) version of this product see product #73367.

Specificity / Sensitivity

Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb detects endogenous HER3/ErbB3 proteins only when phosphorylated at tyrosine 1289. This antibody cross-reacts with overexpressed EGFR. This antibody may cross-react with overexpressed receptor tyrosine kinases in IHC.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Rat, Dog

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1289 of human HER3/ErbB3.

Background

HER3/ErbB3 is a member of the ErbB receptor protein tyrosine kinase family, but it lacks tyrosine kinase activity. Tyrosine phosphorylation of ErbB3 depends on its association with other ErbB tyrosine kinases. Upon ligand binding, heterodimers form between ErbB3 and other ErbB proteins, and ErbB3 is phosphorylated on tyrosine residues by the activated ErbB kinase (1,2). There are at least 9 potential tyrosine phosphorylation sites in the carboxy-terminal tail of ErbB3. These sites serve as consensus binding sites for signal transducing proteins, including Src family members, Grb2, and the p85 subunit of PI3 kinase, which mediate ErbB downstream signaling (3). Both Tyr1222 and Tyr1289 of ErbB3 reside within a YXXM motif and participate in signaling to PI3K (4).

Investigators have found that ErbB3 is highly expressed in many cancer cells (5) and activation of the ErbB3/PI3K pathway is correlated with malignant phenotypes of adenocarcinomas (6). Research studies have demonstrated that in tumor development, ErbB3 may function as an oncogenic unit together with other ErbB members (e.g., ErbB2 requires ErbB3 to drive breast tumor cell proliferation) (7). Thus, investigators view inhibiting interaction between ErbB3 and ErbB tyrosine kinases as a novel strategy for anti-tumor therapy.

  1. Yarden, Y. and Sliwkowski, M.X. (2001) Nat Rev Mol Cell Biol 2, 127-37.
  2. Guy, P.M. et al. (1994) Proc Natl Acad Sci U S A 91, 8132-6.
  3. Songyang, Z. et al. (1993) Cell 72, 767-78.
  4. Kim, H.H. et al. (1994) J Biol Chem 269, 24747-55.
  5. Sithanandam, G. et al. (2003) Carcinogenesis 24, 1581-92.
  6. Kobayashi, M. et al. (2003) Oncogene 22, 1294-301.
  7. Holbro, T. et al. (2003) Proc Natl Acad Sci U S A 100, 8933-8.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 6
#4791

Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb

Western Blotting Image 1: Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb Expand Image
Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb specifically binds to phosphorylated HER3/ErbB3 but not other phosphorylated tyrosine kinases. Western blot analysis of extracts from cells expressing different activated tyrosine kinase proteins, using Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb (upper) or Phospho-Tyrosine mAb (P-Tyr-100) #9411 (lower).
Western Blotting Image 2: Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb Expand Image
Western blot analysis of extracts from mIMCD-3 cells, untreated or treated with neuregulin-1 (100 ng/ml, 5 min) using Phospho-HER3/ErbB3 (Tyr1289) (21D3) RmAb.
Western Blotting Image 1: Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb Expand Image
Simple Western™ analysis of lysates (0.1 mg/mL) MCF-7 +hNRG1 (100ng/mL, 30 mins) cells using Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb #4791. 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.
No image available
Immunohistochemistry Image 1: Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-HER3/ErbB3 (Tyr1289)(21D3) Rabbit mAb.
Immunohistochemistry Image 2: Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb.
Immunohistochemistry Image 3: Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded MDA-MB-453 cells, untreated (left) or neuregulin-treated (right), using Phospho-HER3/ErbB3 (Tyr1289)(21D3) Rabbit mAb.
Immunohistochemistry Image 4: Phospho-HER3/ErbB3 (Tyr1289) (21D3) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, untreated (left) or Calf Intestinal Phosphatase (CIP)-treated (right), using Phospho-HER3/ErbB3 (Tyr1289)(21D3) Rabbit mAb.