bsm-52317R [Primary Antibody]
EGFR Recombinant Antibody
www.biossusa.com
[email protected]
800.501.7654 [DOMESTIC]
+1.781.569.5821 [INTERNATIONAL]
DATASHEET

Host: Rabbit

Target Protein: EGFR

Clonality: Recombinant

Isotype: IgG

Entrez Gene: 1956

Swiss Prot: P00533

Source: Recombinant Human protein.

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol

Storage: Store at 4°C for up to 2 weeks. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.

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.

Size: 100ul

Concentration: 1ug/ul

Predicted Molecular Weight: Predicted: 134kDa; Observed: 150kDa


Cross Reactive Species: Human
Mouse
Rat

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

PRODUCT SPECIFIC PUBLICATIONS
  • Sun Mingyu. et al. Novel molecular typing reveals the risk of recurrence in patients with early-stage papillary thyroid cancer. Thyroid Research. 2024 Dec;17(1):1-12Read more>>
VALIDATION IMAGES

IF(ICC) staining with EGFR (3A8) Monoclonal Antibody (bsm-52317R) at 1:100 in HeLa cells (red). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


IF(ICC) staining with EGFR (3A8) Monoclonal Antibody (bsm-52317R) at 1:100 in A431 cells (red). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


IF(ICC) staining with EGFR (3A8) Monoclonal Antibody (bsm-52317R) at 1:100 in HepG2 cells (red). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


Paraformaldehyde-fixed, paraffin embedded Mouse Skin; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with EGFR Monoclonal Antibody, Unconjugated(bsm-52317R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (EGFR (3A8)) Monoclonal Antibody, Unconjugated (bsm-52317R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.


Paraformaldehyde-fixed, paraffin embedded Rat Skin; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with EGFR Monoclonal Antibody, Unconjugated(bsm-52317R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (EGFR (3A8)) Monoclonal Antibody, Unconjugated (bsm-52317R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.


Paraformaldehyde-fixed, paraffin embedded Rat Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with EGFR Monoclonal Antibody, Unconjugated(bsm-52317R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (EGFR (3A8)) Monoclonal Antibody, Unconjugated (bsm-52317R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.


Paraformaldehyde-fixed, paraffin embedded Rat Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with EGFR Monoclonal Antibody, Unconjugated(bsm-52317R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Lung squamous cell carcinoma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with EGFR Monoclonal Antibody, Unconjugated(bsm-52317R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Lane 1: Human A549 cell lysates; Lane 2: Human U251 cell lysates; Lane 3: Human HeLa cell lysates probed with EGFR Monoclonal Antibody, Unconjugated (bsm-52317R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at room temperature for 60 min.


4% Paraformaldehyde-fixed A549 (left) cell and EGFR knockout A549(right) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (EGFR) monoclonal Antibody, unconjugated (bsm-52317R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-FITC) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei.


Paraformaldehyde-fixed, paraffin embedded Human abdominal skin; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with EGFR Monoclonal Antibody, Unconjugated(bsm-52317R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with EGFR Monoclonal Antibody, Unconjugated(bsm-52317R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


25 ug total protein per lane of various lysates (see on figure) probed with EGFR monoclonal antibody, unconjugated (bsm-52317R) at 1:5000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.