Anti-EGFRvIII antibody (bs-2558R)

Anti-EGFRvIII antibody

DATASHEET

Host:Rabbit

Target Protein:EGFRvIII

IR:Immunogen Range:11-120/1081

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:1956

Swiss Prot:P00533

Source:KLH conjugated synthetic peptide derived from human EGFRvIII:11-120/1081 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:The protein encoded by this gene is a transmembrane glycoprotein that is a member of the protein kinase superfamily. This protein is a receptor for members of the epidermal growth factor family. EGFR is a cell surface protein that binds to epidermal growth factor. Binding of the protein to a ligand induces receptor dimerization and tyrosine autophosphorylation and leads to cell proliferation. Mutations in this gene are associated with lung cancer. Multiple alternatively spliced transcript variants that encode different protein isoforms have been found for this gene. [provided by RefSeq, Jul 2010]

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(3μg/Test)
IF(1:100-500)

Cross Reactive Species:Human
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Protein: A549(human)cell lysates at 40ug;
Primary: rabbit Anti-EGFRIII (bs-2558R) at 1:300; Secondary: HRP conjugated Goat-Anti-rabbit IgG(bs-0295G-HRP) at 1: 5000; Predicted band size:100/130 kD Observed band size:115 kD

Tissue/cell: human laryngocarcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37∩ for 20 min;
Incubation: Anti-EGFRvIII Polyclonal Antibody, Unconjugated(bs-2558R) 1:200, overnight at 4∑C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Tissue/cell: human laryngocarcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37∩ for 20 min;
Incubation: Anti-EGFRvIII Polyclonal Antibody, Unconjugated(bs-2558R) 1:200, overnight at 4∑C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Tissue/cell: SH-SY5Y cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal rabbit serum, C-0006) at 37°C for 20 min; Incubation: Anti-EGFRvIII Polyclonal Antibody, conjugated (bs-2558R-AF647) 1:50, 2 hours at 37°C; DAPI (blue, C02-04002) was used to stain the cell nuclei.

Blank control (blue line): A431 (fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-EGFRvIII antibody (bs-2558R), dilution: 3μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC,Dilution: 1μg /test.

Blank control:SH-SY5Y
Primary Antibody (green line): Rabbit Anti-EGFRvIII antibody (bs-2558R-AF647)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. Acquisition of 20,000 events was performed.