Revision 8

#3197

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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.

Applications:
W, W-S, IP, IHC-P, IF-IC, FC-FP

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
175

Source/Isotype:
Rabbit IgG

UniProt ID:
#P00533

Entrez-Gene Id:
1956

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:250
Immunoprecipitation 1:200
Immunohistochemistry (Paraffin) 1:100
Immunofluorescence (Immunocytochemistry) 1:400
Flow Cytometry (Fixed/Permeabilized) 1:100

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 #28110.

Specificity/Sensitivity

EGFR (L858R Mutant Specific) (43B2) Rabbit mAb detects endogenous levels of EGFR mutant L858R protein. The antibody may cross-react with wildtype EGFR and other HER family members when highly over-expressed. Careful titration of this antibody may be required to obtain optimal specificity.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to L858R mutant sequence of human EGF receptor.

Background

The epidermal growth factor (EGF) receptor is a 170 kDa transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Research studies have shown that somatic mutations in the tyrosine kinase domain of EGF receptor (EGFR) are present in a subset of lung adenocarcinomas that respond to EGFR inhibitors, such as gefitinib and erlotinib (1-3). Two types of mutations account for approximately 90% of mutated cases: a specific point mutation, L858R, that occurs in exon 21 and short in-frame deletions in exon 19 (4,5). The most frequent exon 19 deletion is E746-A750, accounting for 90% of lesions at this site, although some rare variants occur.

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

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: Human

Trademarks and Patents

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

The manufacture, use, sale and import of this product is within the scope of one or more intellectual property rights (including patents and patent applications) owned or controlled by Cell Signaling Technology. The purchase of this product conveys to the buyer a non-transferrable right to use the purchased product only in research conducted by the buyer. The sale of the product is expressly conditioned on the buyer not using the products or its components (1) to analyze or reverse engineer the product for its chemical/physical properties and composition (including e.g., identification of the sequence); (2) in manufacturing; (3) to provide a service, information, or data to an unaffiliated third party for payment; (4) for therapeutic, diagnostic or prophylactic purposes; (5) resale, whether or not such product are resold for use in research; or for any other commercial purpose. For information on purchasing a license to this product for purposes other than research, contact Cell Signaling Technology, Inc. Business Development at [email protected].

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

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除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 , (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

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 8

Cell Signaling Technology Logo
Western blot analysis of extracts from various EGFR expressing cell lines, showing the specificity of detection of mutant EGFR (L858R) protein using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb (upper), and total EGFR control antibody (lower).
Western Blotting Image 1: EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb
Simple Western™ analysis of lysates (0.1 mg/mL) from NCI-H3255 cells using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb #3197. The virtual lane view (left) shows a single target band (as indicated) at a 1:250 dilution of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:250 (grey line) dilution 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.
Western Blotting Image 1: EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded H1975 (left, EGFR L858R positive) and H1650 xenograft (right, EGFR deletion positive) using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb (upper) and total EGFR (D38B1) XP® Rabbit mAb #4267 (lower).
Immunohistochemistry Image 1: EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb
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 8

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human lung carcinoma of known mutational status using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb (upper) and total EGFR (D38B1) XP® Rabbit mAb #4267 (lower). EGFR L858R positive lung (left), EGFR deletion positive lung sections (right).
Immunohistochemistry Image 2: EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb
Confocal immunofluorescent analysis of H3255 (left), HCC827 (middle) and A431 cells (right) using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb (green) and MEK1/2 (L38C12) Mouse mAb #4694 (red). H3255 cells have the L858R EGFR mutation, HCC827 cells have an exon 19 EGFR deletion, and A431 cells overexpress wild type EGFR. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunofluorescence Image 1: EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb
Flow cytometric analysis of two non-small cell lung cancer cell lines using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb. H3255 cells (green) harbor the L858R EGFR mutation while the H1650 cells (blue) have the exon 19 mutation (delE746-A750).
Flow Cytometry Image 1: EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb
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.