EGFR Monoclonal antibody

EGFR Monoclonal Antibody for WB, IF/ICC, FC, ELISA, Blocking

Host / Isotype

Mouse / IgG1

Reactivity

human

Applications

WB, IF/ICC, FC, ELISA, Blocking

Conjugate

Unconjugated

CloneNo.

2C8C12

Cat no : 68643-1-Ig

Synonyms

Proto-oncogene c-ErbB-1, Epidermal growth factor receptor, EGF Receptor , EGF R, 2C8C12

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Validation Data Gallery

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  WB analysis of SCaBER using 68643-1-Ig

  SCaBER cells were subjected to SDS PAGE followed by western blot with 68643-1-Ig (EGFR antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of MDA-MB-468 using 68643-1-Ig

  MDA-MB-468 cells were subjected to SDS PAGE followed by western blot with 68643-1-Ig (EGFR antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 68643-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68643-1-Ig (EGFR antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  IF Staining of HaCaT using 68643-1-Ig

  Immunofluorescent analysis of un-fixed HaCaT cells using EGFR antibody (68643-1-Ig, Clone: 2C8C12 ) at dilution of 1:1000 and Multi-rAb CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM004 ).

• 

  IF Staining of A431 using 68643-1-Ig

  Immunofluorescent analysis of un-fixed A431 cells using EGFR antibody (68643-1-Ig, Clone: 2C8C12 ) at dilution of 1:1000 and Multi-rAb CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM004 ).

• 

  FC experiment of A431 using 68643-1-Ig

  1x10^6 A431 cells were surface stained with 0.2 μg EGFR Monoclonal Antibody (68643-1-Ig, Clone:2C8C12) and CoraLite488-conjugated Goat Anti-Mouse IgG(H+L) (Cat.NO. SA00013-1)(red), or 0.2 μg Mouse IgG1 Isotype Control (66360-1-Ig, Clone: T1F8D3F10) (blue). Cells were not fixed.

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  Blocking experiment of HeLa cells using 68643-1-Ig

  Human EGF (Cat.NO. HZ-1326 ) stimulates proliferation of HeLa cells in a dose-dependent manner (blue curve, refer to bottom X-left Y axis). The activity of human EGF (Cat.NO. HZ-1326 ) is blocked by mouse anti-human EGFR monoclonal antibody 68643-1-Ig at serial dose (red curve, refer to top X-right Y axis). The EC50 is typically 0.2-0.8 μg/mL at the presence of 1ng/ml EGF (Cat.NO. HZ-1326 ).

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Tested Applications

• Positive WB detected in: SCaBER cells, HaCaT cells, MDA-MB-468 cells, A431 cells
• Positive IF/ICC detected in: HaCaT cells, A431 cells
• Positive FC detected in: A431 cells
• Positive Blocking detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:500-1:2000
• Flow Cytometry (FC): FC : 0.20 ug per 10^6 cells in a 100 µl suspension
• BLOCKING: BLOCKING : 1:10-1:100

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

68643-1-Ig targets EGFR in WB, IF/ICC, FC, ELISA, Blocking applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: EGFR fusion protein Ag24947
• Full Name: epidermal growth factor receptor (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian)
• Calculated Molecular Weight: 134kd
• Observed Molecular Weight: 160 kDa
• GenBank Accession Number: NM_005228.5
• Gene Symbol: EGFR
• Gene ID (NCBI): 1956
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

EGFR, also named ERBB1, is a cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein ligands. Binding of the protein to a ligand induces receptor dimerization and tyrosine autophosphorylation and leads to cell proliferation. The gene resides on chromosome 7p12, encoding a 170 kDa membrane-associated glycoprotein. Recent studies have shown EGFR plays a critical role in cancer development and progression, including cell proliferation, apoptosis, angiogenesis, and metastatic spread. Mutations in this gene are associated with lung cancer. When stained with live cells, EGFR can be transported into the cell interior, which is consistent with the literature report (PMID: 18084617).