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EGFR Polyclonal Antibody

Formalin-fixed and paraffin-embedded human lung carcinoma labeled with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Formalin-fixed and paraffin-embedded human lung carcinoma labeled with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Paraformaldehyde-fixed, paraffin embedded human brain glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:400 overnight at 4\u00b0C, followed by a conjugated secondary for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded human brain glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:400 overnight at 4\u00b0C, followed by a conjugated secondary for 20 minutes and DAB staining.
Tissue\/cell: Human Rectal Carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15min; Blocking buffer (normal goat serum) at 37\u2103 for 20 min; Incubation: EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200, overnight at 4\u00b0C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (bs-0295G-Cy3) used at 1:200 dilution for 40 minutes at 37\u00b0C. DAPI(5ug\/ml, blue) was used to stain the cell nuclei.
Tissue\/cell: Human Rectal Carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15min; Blocking buffer (normal goat serum) at 37\u2103 for 20 min; Incubation: EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200, overnight at 4\u00b0C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (bs-0295G-Cy3) used at 1:200 dilution for 40 minutes at 37\u00b0C. DAPI(5ug\/ml, blue) was used to stain the cell nuclei.
Human HUVEC Cells were fixed with 2% PFA (10min at room temperature) and then permeabilized with 90% ice-cold methanol for 30min on ice. The cells were then incubated in 0.5% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min. Cells were stained with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:100 for 30 min on ice. The secondary antibody, Goat anti-rabbit IgG-PE, was used for 30 min on ice. Primary Antibody staining (green) is compared to unstained cells (dark blue), secondary only (light blue) and isotype control (orange).
Human HUVEC Cells were fixed with 2% PFA (10min at room temperature) and then permeabilized with 90% ice-cold methanol for 30min on ice. The cells were then incubated in 0.5% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min. Cells were stained with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:100 for 30 min on ice. The secondary antibody, Goat anti-rabbit IgG-PE, was used for 30 min on ice. Primary Antibody staining (green) is compared to unstained cells (dark blue), secondary only (light blue) and isotype control (orange).
Human A431 Cells were fixed with 2% PFA (10min at room temperature) and then permeabilized with 90% ice-cold methanol for 30min on ice. The cells were then incubated in 2% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min at room temperature. Cells were stained with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:33 for 30 min at room temperature. The secondary antibody, Goat anti-rabbit IgG-FITC, was used for 40 min at room temperature. Primary Antibody staining (green) is compared to unstained cells (dark blue), secondary only (light blue) and isotype control (orange).
Human A431 Cells were fixed with 2% PFA (10min at room temperature) and then permeabilized with 90% ice-cold methanol for 30min on ice. The cells were then incubated in 2% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min at room temperature. Cells were stained with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:33 for 30 min at room temperature. The secondary antibody, Goat anti-rabbit IgG-FITC, was used for 40 min at room temperature. Primary Antibody staining (green) is compared to unstained cells (dark blue), secondary only (light blue) and isotype control (orange).
Paraformaldehyde-fixed, paraffin embedded (rat placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (EGFR) Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (EGFR) Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (EGFR) Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (EGFR) Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Applications

  • WB
  • ELISA
  • FCM
  • IHC-P
  • IHC-F
  • IF(IHC-P)
  • IF(IHC-F)
  • IF(ICC)

Reactivity

  • Human

Predicted Reactivity

  • Mouse
  • Rat
  • Dog
  • Pig
Overview
Catalog # bs-0165R
Product Name EGFR Polyclonal Antibody
Applications WB, ELISA, FCM, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC)
Reactivity Human
Predicted Reactivity Mouse, Rat, Dog, Pig
Specifications
Conjugation Unconjugated
Host Rabbit
Source KLH conjugated synthetic peptide derived from human EGFR
Immunogen Range 951-1050/1210
Clonality Polyclonal
Isotype IgG
Concentration 1ug/ul
Purification Purified by Protein A.
Storage Buffer 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID 1956
Swiss Prot P00533
Subcellular location Cytoplasm, Nucleus, Secreted, Cell membrane
Synonyms ERBB; HER1; mENA; ERBB1; PIG61; NISBD2; Epidermal growth factor receptor; Proto-oncogene c-ErbB-1; Receptor tyrosine-protein kinase erbB-1; EGFR
Background Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses. Known ligands include EGF, TGFA/TGF-alpha, amphiregulin, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. Ligand binding triggers receptor homo- and/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. Activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules. May also activate the NF-kappa-B signaling cascade. Also directly phosphorylates other proteins like RGS16, activating its GTPase activity and probably coupling the EGF receptor signaling to the G protein-coupled receptor signaling. Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin. Isoform 2 may act as an antagonist of EGF action.
Application Dilution
WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

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