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05-104

Sigma-Aldrich

Anti-EGFR Antibody, clone LA22

clone LA22, Upstate®, from mouse

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Synonym(s):
Receptor tyrosine-protein kinase ErbB-1, avian erythroblastic leukemia viral (v-erb-b) oncogene homolog, cell growth inhibiting protein 40, cell proliferation-inducing protein 61, epidermal growth factor receptor, epidermal growth factor receptor (avian
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

clone

LA22, monoclonal

species reactivity

human

manufacturer/tradename

Upstate®

technique(s)

immunocytochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

isotype

IgG2a

NCBI accession no.

UniProt accession no.

shipped in

dry ice

target post-translational modification

unmodified

Gene Information

human ... EGFR(1956)

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antibody form

purified antibody

antibody form

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antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

clone

LA22, monoclonal

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8G6.2, monoclonal

clone

7F12.1, monoclonal

clone

3E7.2, monoclonal

species reactivity

human

species reactivity

rat, mouse, human

species reactivity

human

species reactivity

human

manufacturer/tradename

Upstate®

manufacturer/tradename

-

manufacturer/tradename

-

manufacturer/tradename

-

technique(s)

immunocytochemistry: suitable, immunoprecipitation (IP): suitable, western blot: suitable

technique(s)

immunocytochemistry: suitable, immunohistochemistry: suitable, immunoprecipitation (IP): suitable, western blot: 1:2,000 using A431 cell lysate (used to detect EGFR), western blot: suitable

technique(s)

flow cytometry: suitable, immunocytochemistry: suitable, western blot: suitable

technique(s)

immunocytochemistry: suitable, western blot: suitable

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General description

Protein kinases are enzymes that transfer a phosphate group from a phosphate donor onto an acceptor amino acid in a substrate protein. By this basic mechanism, protein kinases mediate most of the signal transduction in eukaryotic cells, regulating cellular metabolism, transcription, cell cycle progression, cytoskeletal rearrangement and cell movement, apoptosis, and differentiation. The protein kinase family is one of the largest families of proteins in eukaryotes, classified in 8 major groups based on sequence comparison of their tyrosine (PTK) or serine/threonine (STK) kinase catalytic domains. Epidermal Growth factor receptor (EGFR) is the prototype member of the type 1 receptor tyrosine kinases. EGFR overexpression in tumors indicates poor prognosis and is observed in tumors of the head and neck, brain, bladder, stomach, breast, lung, endometrium, cervix, vulva, ovary, esophagus, stomach and in squamous cell carcinoma.

Specificity

Recognizes the sequence of Ala351-Asp364 of human EGF receptor, Mr 180kDa, (based on numbering without the signal peptide) (Wu et al., 1989).

Immunogen

A431 membrane proteins

Application

Anti-EGFR Antibody, clone LA22 is an antibody against EGFR for use in IC, IP & WB.
Immunoprecipitation:
4 μg of a previous lot immunoprecipitated EGF receptor from 500 μg of A431 RIPA lysate.

Immunocytochemistry:
5 μg/mL of previous lots of the antibody showed positive immunostaining for EGF receptor in A431 cells fixed with 4% paraformaldehyde followed by 100% methanol.
Research Category
Signaling
Research Sub Category
Growth Factors & Receptors

Quality

Evaluatd by RIPA lysates from human A431 cells under reducing conditions.

Western Blot Analysis:
0.5-2 μg/mL of this antibody detected EGF receptor in RIPA lysates from human A431 cells under reducing conditions.

Target description

180 kDa

Linkage

Replaces: 04-337; 04-338

Physical form

Format: Purified
Protein G Purified
Purified mouse monoclonal IgG2a in storage buffer (0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide). Frozen solution.

Storage and Stability

Stable for 1 year at -20ºC from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Analysis Note

Control

Positive Antigen Control: Catalog #12-301, non-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for minigels.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

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Leif D Nelin et al.
American journal of respiratory cell and molecular biology, 33(4), 394-401 (2005-07-05)
L-arginine is metabolized to nitric oxide (NO) by NO synthase (NOS), or to urea and L-ornithine by arginase. L-ornithine contributes to vascular remodeling in pulmonary hypertension via metabolism to polyamines and proline. Previously we found that cytokines upregulate both NOS
SHC and GRB-2 are constitutively by an epidermal growth factor receptor with a point mutation in the transmembrane domain.
Miloso, M, et al.
The Journal of Biological Chemistry, 270, 19557-19562 (1995)
Endocytosis of epidermal growth factor receptor regulated by Grb2-mediated recruitment of the Rab5 GTPase-activating protein RN-tre.
Martinu, L; Santiago-Walker, A; Qi, H; Chou, MM
The Journal of Biological Chemistry null
Jillian M Prendergast et al.
mAbs, 9(4), 615-627 (2017-03-11)
Targeted therapeutics that can differentiate between normal and malignant tumor cells represent the ideal standard for the development of a successful anti-cancer strategy. The Sialyl-Thomsen-nouveau antigen (STn or Sialyl-Tn, also known as CD175s) is rarely seen in normal adult tissues
Monoclonal antibodies to epidermal growth factor receptors in studies of receptor structure and function.
Kawamoto, T, et al.
Cytotechnology, 3, 279-293 (1990)

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