PRODUCT CODE: EM1901-67

EGFR Mouse Monoclonal Antibody [7-F2-F8] (EM1901-67)

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

Western blot analysis of EGFR on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-67, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: A431 cell lysate<br />
Lane 2: SKOV-3 cell lysate<br />
Lane 3: A549 cell lysate
  • Western blot analysis of EGFR on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-67, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: A431 cell lysate<br />
Lane 2: SKOV-3 cell lysate<br />
Lane 3: A549 cell lysate
  • ICC staining of EGFR in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1901-67, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-EGFR antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-67, 1/50)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-EGFR antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-67, 1/50)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of EGFR was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-67, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of EGFR on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-67, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: A431 cell lysate
Lane 2: SKOV-3 cell lysate
Lane 3: A549 cell lysate

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

EGFR Mouse Monoclonal Antibody [7-F2-F8] (EM1901-67)

Immunogen

Recombinant protein within human egfr aa 900-1150.

Host

Mouse

Positive Control

A431 cell lysate, SKOV-3 cell lysate, A549 cell lysate, A549, human breast carcinoma tissue, human placenta tissue, A431.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

7-F2-F8

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

Predicted band size 134 kDa.

Isotype

IgG2b

APPLICATION DILUTION

  • WB:1:500-1:2,000

  • ICC:1:50-1:100

  • IHC-P:1:50-1:200

  • FC:1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

EGFR

SYNONYMS

Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody; Cell growth inhibiting protein 40 antibody; Cell proliferation inducing protein 61 antibody; EGF R antibody; EGFR antibody; EGFR_HUMAN antibody; Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog) antibody; Epidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian) antibody; Epidermal growth factor receptor antibody; erb-b2 receptor tyrosine kinase 1 antibody; ERBB antibody; ERBB1 antibody; Errp antibody; HER1 antibody; mENA antibody; NISBD2 antibody; Oncogen ERBB antibody; PIG61 antibody; Proto-oncogene c-ErbB-1 antibody; Receptor tyrosine protein kinase ErbB 1 antibody; Receptor tyrosine-protein kinase ErbB-1 antibody; SA7 antibody; Species antigen 7 antibody; Urogastrone antibody; v-erb-b Avian erythroblastic leukemia viral oncogen homolog antibody; wa2 antibody; Wa5 antibody

SEQUENCE SIMILARITIES

Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.

TISSUE SPECIFICITY

Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated on Tyr residues in response to EGF. Phosphorylation at Ser-695 is partial and occurs only if Thr-693 is phosphorylated. Phosphorylation at Thr-678 and Thr-693 by PRKD1 inhibits EGF-induced MAPK8/JNK1 activation. Dephosphorylation by PTPRJ prevents endocytosis and stabilizes the receptor at the plasma membrane. Autophosphorylation at Tyr-1197 is stimulated by methylation at Arg-1199 and enhances interaction with PTPN6. Autophosphorylation at Tyr-1092 and/or Tyr-1110 recruits STAT3. Dephosphorylated by PTPN1 and PTPN2.; Monoubiquitinated and polyubiquitinated upon EGF stimulation; which does not affect tyrosine kinase activity or signaling capacity but may play a role in lysosomal targeting. Polyubiquitin linkage is mainly through 'Lys-63', but linkage through 'Lys-48', 'Lys-11' and 'Lys-29' also occurs. Deubiquitination by OTUD7B prevents degradation. Ubiquitinated by RNF115 and RNF126 (By similarity).; Palmitoylated on Cys residues by ZDHHC20. Palmitoylation inhibits internalization after ligand binding, and increases the persistence of tyrosine-phosphorylated EGFR at the cell membrane. Palmitoylation increases the amplitude and duration of EGFR signaling.; Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197.

SUBCELLULAR LOCATION

Golgi apparatus membrane, nucleus membrane, nucleus, cell membrane, endosome, endosome membrane, endoplasmic reticulum membrane.

FUNCTION

The EGF receptor family comprises several related receptor tyrosine kinases that are frequently overexpressed in a variety of carcinomas. Members of this receptor family include EGFR (HER1), Neu (ErbB-2, HER2), ErbB-3 (HER3) and ErbB-4 (HER4), which form either homodimers or heterodimers upon ligand binding. EGFR binds several ligands, including epidermal growth factor (EGF), transforming growth factor α (TGFα), Amphiregulin and heparin binding-EGF (HB-EGF). Ligand binding promotes the internalization of EGFR via Clathrin-coated pits and its subsequent degradation in response to its intrinsic tyrosine kinase. EGFR is involved in organ morphogenesis and maintenance and repair of tissues, but upregulation of EGFR is associated with tumor progression. The oncogenic effects of EGFR include initiation of DNA synthesis, enhanced cell growth, invasion and metastasis. Abrogation of EGFR results in cell cycle arrest, apoptosis or dedifferentiation of cancer cells, suggesting that EGFR may be an effective therapeutic target.