WB result of EGFR (L858R) Recombinant Rabbit mAb
Primary antibody: EGFR (L858R) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: A431 whole cell lysate 20 µg
Lane 3: A549 whole cell lysate 20 µg
Lane 4: HCC827 whole cell lysate 20 µg
Lane 5: H1975 whole cell lysate 20 µg
Negative control: HeLa whole cell lysate; A431 whole cell lysate; A549 whole cell lysate; HCC827 whole cell lysate
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 134 kDa
Observed MW: 150 kDa
EGFR (L858R) Recombinant Rabbit mAb (SDT-421-202)
EGFR (L858R) Recombinant Rabbit mAb (SDT-421-202)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | EGFR (L858R) |
| Synonyms | Epidermal growth factor receptor; Proto-oncogene c-ErbB-1; Receptor tyrosine-protein kinase erbB-1; ERBB; ERBB1; HER1 |
| Immunogen | Synthetic Peptide |
| Location | Cell membrane, Endoplasmic reticulum membrane |
| Accession | P00533 |
| Clone Number | SDT-421-202 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, IP |
| Reactivity | Hu |
| Positive Sample | H1975 |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| Dot Blot | 1:1000 | |
| WB | 1:1000 | Hu |
| IP | 1:50 | Hu |
| IHC-P | 1:100-1:200 | Hu |
Background
EGFR is a 170 kDa transmembrane receptor that induces tyrosine kinase activity affecting cell growth. EGFR mutations can occur in various parts of the gene. In NSCLC, the mutations occur almost exclusively in the exons encoding the kinase domain (exons 18 to 21). They most commonly include the in-frame deletion of exon 19 (del19) and L858R mutation in exon 21.Del19 constitutes 44–51%, and exon 21 L858R mutation constitutes 38–40%.
Picture
Picture
Western Blot
IP
EGFR(L858R) Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating EGFR(L858R) in 0.4 mg H1975 whole cell lysate.
Western blot was performed on the immunoprecipitate using EGFR(L858R) Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: H1975 whole cell lysate 20 µg (Input)
Lane 2: EGFR(L858R) Rabbit mAb IP in H1975 whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in H1975 whole cell lysate
Predicted MW: 134 kDa
Observed MW: 175 kDa
Dot Blot
Dot blot result of EGFR (L858R) Recombinant Rabbit mAb
Lane1: EGFR(L858R) mutated peptide
Lane2: EGFR(L858R) mutated peptide
Lane3: EGFR unmutated peptide
Primary antibody: EGFR (L858R) Recombinant Rabbit mAb at 1/1000 dilution
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human lung adenocarcinoma (Mutated case). Anti-EGFR (L858R) antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human lung adenocarcinoma (un-mutated case). Anti-EGFR (L858R) antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human lung. Anti-EGFR (L858R) antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
