Active Recombinant Human EGFR protein, His-tagged, Low Endotoxin

• Cat.No.: EGFR-628H
• Product Overview: Recombinant Human EGFR protein(NP_001333870.1)(Leu25-Ser378), fused with C-terminal His tag, was expressed in HEK293.

Specification

• Species: Human
• Source: HEK293
• Tag: His
• Protein Length: 25-378 aa
• Form: Lyophilized from 0.22 μm filtered solution in PBS (pH 7.4). Normally 8% trehalose is added as protectant before lyophilization.
• Bio-activity: Immobilized Human EGFRVIII, His Tag at 1μg/ml (100μl/well) on the plate. Dose response curve for Anti-EGFRVIII Antibody, hFc Tag with the EC50 of 43.3ng/ml determined by ELISA.
• Molecular Mass: The protein has a predicted MW of 40.25 kDa. Due to glycosylation, the protein migrates to 60-80 kDa based on Bis-Tris PAGE result.
• Endotoxin: Less than 0.01 EU per μg by the LAL method.
• Purity: > 95% as determined by Bis-Tris PAGE; > 95% as determined by HPLC
• Storage: Store at -20 to -80°C for 24 months as supplied from date of receipt. -80°C for 3 months after reconstitution. Recommend to aliquot the protein into smaller quantities for optimal storage. Please minimize freeze-thaw cycles.
• Reconstitution: Dissolve the lyophilized protein in distilled water.

Gene Information

• Gene Name: EGFR epidermal growth factor receptor [ Homo sapiens ]
• Official Symbol: EGFR
• Synonyms: EGFR; epidermal growth factor receptor; epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog), ERBB; ERBB1; erythroblastic leukemia viral (v erb b) oncogene homolog (avian); proto-oncogene c-ErbB-1; cell growth inhibiting protein 40; cell proliferation-inducing protein 61; receptor tyrosine-protein kinase erbB-1; avian erythroblastic leukemia viral (v-erb-b) oncogene homolog; ERBB; HER1; mENA; PIG61
• Gene ID: 1956
• mRNA Refseq: NM_005228
• Protein Refseq: NP_005219
• MIM: 131550
• UniProt ID: P00533

Citation

Regulation of EGFR Protein Stability by the HECT-type Ubiquitin Ligase SMURF2

Journal: Neoplasia (New York, N.Y.)    PubMed ID: 21750651    Data: 2011/7/1

Authors: Dipankar Ray, Aarif Ahsan, Mukesh K Nyati

Article Snippet:GST-SMURF2 was purified from bacteria as previously described [ 30 ], and 1 μg of fusion protein still attached to agarose beads was equilibrated in 0.5x Superdex buffer (1x Superdex buffer: 25 mM HEPES, pH 7.5, 12.5 mM MgCl 2 , 10 μM ZnSO 4 , 150 mM KCl, 20% glycerol, 0.1% Nonidet P-40, and 1 mM EDTA) for 30 minutes at 4°C and then washed three times with 0.5x Superdex buffer.agarose beads was equilibrated in 0.5x Superdex buffer (1x Superdex buffer: 25 mM HEPES, pH 7.5, 12.5 mM MgCl 2 , 10 μM ZnSO 4 , 150 mM KCl, 20% glycerol, 0.1% Nonidet P-40, and 1 mM EDTA) for 30 minutes at 4°C and then washed three times with 0.5x Superdex buffer. ... Beads were then incubated with either about 200 or 600 ng of purified His-tagged EGFR protein (Creative BioMart, New York, NY) was then added to the washed beads and incubated overnight at 4°C.. The beads were washed three times using 0.5 Superdex buffer and boiled in Laemmli buffer, and the bound SMURF2-EGFR complex was immunodetected after immunoblot analysis with SMURF2 and Tetra-His (for EGFR) antibodies.The beads were washed three times using 0.5 Superdex buffer and boiled in Laemmli buffer, and the bound SMURF2-EGFR complex was immunodetected after immunoblot analysis with SMURF2 and Tetra-His (for EGFR) antibodies.