BD Pharmingen™

Technical Data Sheet

Alexa Fluor® 647 Mouse Anti-Human EGF Receptor

Product Information

Material Number: 563577

Alternate Name: EGFR; ERBB; ERBB1; HER1; mENA; PIG61

Size: 50 tests

Vol. per Test: 5 µl

Clone: EGFR.1

Isotype: Mouse IgG2b, κ

QC Testing: HumanReactivity:

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

DescriptionThe EGFR.1 monoclonal antibody specifically binds to the epidermal growth factor receptor (EGF-R). The EGF-R is a transmembrane

glycoprotein of approximately 170 kDa that is expressed on most cells. It consists of a glycosylated extracellular domain which binds to

epidermal growth factor (EGF), a transmembrane domain, and an intracellular domain with tyrosine-kinase activity essential for signal

transduction. The EGF-R plays an important role in the growth and differentiation of many cellular types. Transforming growth factor α

(TGFα), vaccinia virus growth factor, and related growth factors can also bind to and signal through the EGF-R.

Flow cytometric analysis of EGF Receptor expression

on human A-431 cells. Cells from the human A-431

epidermoid carcinoma cell Line (ATCC CRL-1555) were

stained with Alexa Fluor® 647 Mouse Anti-Human EGF

Receptor antibody (Cat. No. 563577; solid line histogram) or

with Alexa Fluor® 647 Mouse IgG2b, κ Isotype Control (Cat.

No. 557903; dashed line histogram). The fluorescence

histograms were derived from gated events with the forward

and side light-scatter characteristics of viable cells. Flow

cytometric analysis was performed using a BD™ LSR II

Flow Cytometer System.

Preparation and StorageStore undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Application Notes

Application

Flow cytometry Routinely Tested

Suggested Companion Products

Catalog Number Name CloneSize

554656 Stain Buffer (FBS) 500 ml (none)

557903 Alexa Fluor® 647 Mouse IgG2b, κ Isotype Control 100 tests 27-35

563577 Rev. 1 Page 1 of 2

Product NoticesThis reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental

sample (a test).

1.

Source of all serum proteins is from USDA inspected abattoirs located in the United States. 2.

An isotype control should be used at the same concentration as the antibody of interest. 3.

Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols. 4.

The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular

Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes

(except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.

5.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR. 6.

Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC). 7.

Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before

discarding to avoid accumulation of potentially explosive deposits in plumbing.

8.

For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at

www.bdbiosciences.com/colors.

9.

ReferencesFernandez A, Spitzer E, Perez R. A new monoclonal antibody for detection of EGF-receptors in western blots and paraffin-embedded tissue sections. J Cell Biol.

1992; 49(2):157-165. (Biology)

Fitzgerald KA, O Neill LAJ, Geraring AJH, Callard, RE. The Cytokine FactsBook. San Diego, CA: Academic Press; 2001:1-515. (Biology)

Mishima K, Johns TG, Luwor RB, Scott AM, Stockert E, Jungbluth AA, Ji XD, Suvarna P, Voland JR, Old LJ, Huang HJ, Cavenee WK. Growth suppression of

intracranial xenografted glioblastomas overexpressing mutant epidermal growth factor receptors by systemic administration of monoclonal antibody (mAb) 806, a

novel monoclonal antibody directed to the receptor. Cancer Res. 2001; 61(14):5349-5354. (Clone-specific: Flow cytometry, Immunoprecipitation)

Troalen F, Maudelondé T, Montcourrier P, Bohuon C, Delarue JC. Characterization of two monoclonal antibodies against the COOH-terminal part of the human

epidermal growth factor receptor and potential clinical use. J Immunol Methods. 1991; 143(2):175-186. (Biology)

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