Antibodies to the Epidermal Growth Factor Receptor-Family EGFR and HER2,-3,-4 - Product Review 13

Introduction

The epidermal growth factor (EGF) family of receptor tyrosine kinases consists of four receptors, EGF-R (HER1, ERBB1), HER2 (neu) (ERBB2, c-erbB-2), HER3 (ERBB3, c-erbB-3), and HER4 (ERBB4, p180erbB4).

Members of the EGF-R family contain a cytoplasmic tyrosine kinase domain, a single transmembrane domain, and an extracellular domain that is involved in ligand binding and receptor dimerization. Activation of the EGF-R results in the initiation of a diverse array of cellular pathways. In response to toxic environmental stimuli, such as ultraviolet irradiation, or to receptor occupation by EGF, the EGF-R forms homo- or heterodimers with other family members. Each dimeric receptor complex will initiate a distinct signaling pathway by recruiting different Src homology 2 (SH2)-containing effector proteins. Dimerization results in autophosphorylation initiating a downstream cascade of events culminating in cellular responses such as cell proliferation or apoptosis. Therefore, overactivation of epidermal growth factor receptor (EGFR) signaling has been recognized as an important step in the pathogenesis and progression of multiple forms of cancer of epithelial origin:

The overexpression of HER-1 and HER-2 have been reported previously in surface epithelial ovarian cancer. HER-2 measurement is carried out in primary breast tumors as its overexpression occurs in 20-30 % of breast cancer and is a predictive marker of tumor aggressiveness and responsiveness to therapy. For metaplastic carcinomas (MC) of the breast it has been shown recently that they rarely express steroid receptors and HER-2: MCs express EGFR considerably more frequently.

While HER-1 (EGF-receptor) and HER-2 have been extensively studied, little is known about the clinical significance of HER-3 and especially HER-4 expression. For instance, the expression of HER-3 and HER-4 has been investigated in pancreatic cancer and osteaosarcoma. The overexpression of HER-3 protein has also been shown in 53%-88% of colorectal adenocarcinomas.

Antibody Tools for Detection of EGF-Receptor and HER-2,-3,-4

Antibody products for the detection of EGF-R including a range of phospho-specific antibodies (and their corresponding control peptides) are available at Acris Antibodies. All antibodies are thoroughly tested for various applications, many of them can be used for immunohistochemistry and nearly all are suitable for western blotting.

Picture Gallery

Western blot image of EGFR from human HeLa cells using antibody SP5432PImmunofluorescence image of EGFR in HeLa cells using antibody SP5432PFormalin-fixed, paraffin-embedded human lung squamous cell carcinoma: membrane staining  of tumor cells with EGFR antibody DM267 using peroxidase-conjugate and AEC chromogen.

Fig.1: Western blot image of EGFR from human HeLa cells using antibody SP5432P

Fig.2: Immunofluorescence image of EGFR in HeLa cells using antibody SP5432P

Fig.3: Formalin-fixed, paraffin-embedded human lung squamous cell carcinoma: membrane staining of tumor cells with EGFR antibody DM267 using peroxidase-conjugate and AEC chromogen.

Formalin-fixed, paraffin-embedded human breast carcinoma: intense membrane staining of tumor cells with c-erbB-2 antibody BM5084 using peroxidase-conjugate and DAB chromogen.Anti-HER-2 IgY: Immunohistochemical staining of Formalin-fixed and Paraffin-embedded esophageal cancer tissue using anti-HER-2 IgY antibody 15-288-22631 (Affi-pure IgY dilution 1:100, Rabbit anti IgY-Biotin-dilution 1:300Negative control: Immunohistochemical staining of Formalin-fixed and Paraffin-embedded esophageal cancer tissue using anti-HER-2 IgY antibody 15-288-22631 (Affi-pure IgY dilution 1:100, Rabbit anti IgY-Biotin-dilution 1:300

Fig.4: Formalin-fixed, paraffin-embedded human breast carcinoma: intense membrane staining of tumor cells with c-erbB-2 antibody BM5084 using peroxidase-conjugate and DAB chromogen.

Fig.5 ( Anti-HER-2 IgY ) and 6 (Negative control): Immunohistochemical staining of Formalin-fixed and Paraffin-embedded esophageal cancer tissue using anti-HER-2 IgY antibody 15-288-22631 (Affi-pure IgY dilution 1:100, Rabbit anti IgY-Biotin-dilution 1:300 (antibody manufactured by GenWay Biotech, Inc., pictures courtesy of Jessica Wang-Rodriguez MD, San Diego Healthcare System)

Immunocytochemical staining of HER-2 in human breast cancer cell line SKBR3 using antibody 15-288-22631 (IgY dilution 1:1000, Goat anti IgY Fc-FITC 1:200)Formalin-fixed, paraffin-embedded human skeletal muscle stained with c-erbB-3 antibody SM1444P using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of muscle cells.Formalin-fixed, paraffin-embedded human breast cancer: tumor cell membrane staining with c-erbB-4 antibody SM5154P using peroxidase-conjugate and AEC chromogen.

Fig.7: Immunocytochemical staining of HER-2 in human breast cancer cell line SKBR3 using antibody 15-288-22631 (IgY dilution 1:1000, Goat anti IgY Fc-FITC 1:200) (antibody manufactured by GenWay Biotech, Inc., data provided by Yan Xiao & Peter E. Barker, NIST)

Fig.8: Formalin-fixed, paraffin-embedded human skeletal muscle stained with c-erbB-3 antibody SM1444P using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of muscle cells.

Fig.9: Formalin-fixed, paraffin-embedded human breast cancer: tumor cell membrane staining with c-erbB-4 antibody SM5154P using peroxidase-conjugate and AEC chromogen.

Available Antibodies to the EGF Receptor-Family

Anti- EGFR (ERBB1), c-erbB-2 ( HER2, ERBB2), c-erbB-3 (HER3, ERBB3), c-erbB-4 (HER4) antibody

Literature

Valentini AM, Pirelli M, Caruso ML.EGFR-targeted therapy in colorectal cancer: does immunohistochemistry deserve a role in predicting the response to cetuximab? Curr Opin Mol Ther. 2008 Apr;10(2):124-31

Bodey B, Kaiser HE, Siegel SE.Epidermal growth factor receptor (EGFR) expression in childhood brain tumors. In Vivo. 2005 Sep-Oct; 19(5): 931-41

Zidan J, Dashkovsky I, Stayerman C, Basher W, Cozacov C, Hadary A.Comparison of HER-2 overexpression in primary breast cancer and metastatic sites and its effect on biological targeting therapy of metastatic disease.Br J Cancer. 2005 Sep 5; 93(5): 552-6

Leibl S, Moinfar F.Metaplastic breast carcinomas are negative for Her-2 but frequently express EGFR (Her-1): potential relevance to adjuvant treatment with EGFR tyrosine kinase inhibitors ?J Clin Pathol. 2005 Jul; 58(7): 700-4

Lee CH, Huntsman DG, Cheang MC, Parker RL, Brown L, Hoskins P, Miller D, Gilks CB.Assesment of Her-1, Her-2, and Her-3 expression and Her-2 amplification in advanced stage ovarian carcinoma.Int J Gynecol Pathol. 2005 Apr; 24(2): 147-52

Hughes DP, Thomas DG, Giordano TJ, Baker LH, McDonagh KT.Cell surface expression of epidermal growth factor receptor and Her-2 with nuclear expression of Her-4 in primary osteosarcoma.Cancer Res. 2004 Mar 15; 64(6): 2047-53

Thybusch-Bernhardt A, Beckmann S, Juhl M.Comparative analysis of the EGF-receptor family in pancreatic cancer: expression of HER-4 correlates with a favourable tumor stage.Int J Surg Investig. 2001; 2(5): 393-400

Kapitanovic S, Radosevic S, Slade N, Kapitanovic M, Andelinovic S, Ferencic Z, Tavassoli M, Spaventi S, Pavelic K, Spaventi R.Expression of erbB-3 protein in colorectal adenocarcinoma: correlation with poor survival.J Cancer Res Clin Oncol. 2000 Apr; 126(4): 205-11

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

ERBB3 / HER3 pTyr1328 antibody

Figure 1. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Her3/ErbB3 pTyr1328 Antibody (#AP08032PU). Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, P, WB
0.1 ml / €320.00
  Acris Antibodies GmbH

ERBB3 / HER3 pTyr1328 antibody

Figure 1. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Her3/ErbB3 pTyr1328 Antibody (#AP08032PU). Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, P, WB
50 µl / €240.00
  Acris Antibodies GmbH

ERBB3 / HER3 pTyr1328 antibody

Immunohistochemistry: Her3/ErbB3 phosphospecific Antibody [NBP1-04970] - Immunohistochemical analysis of paraffin-embedded  human breast carcinoma tissue using Her3/ErbB3  (phospho-Tyr1328) Antibody Rabbit IgG Aff - Purified Hu, Ms, Rt ICC/IF, P, WB
0.1 mg / €420.00
  Novus Biologicals Inc.

ERBB3 / HER3 antibody

AM09108PU c-erbB3 antibody staining of Human Small Intestine. Rabbit IgG SP71 Purified Hu P

Regular Price: 1 ml / €730.00

Special Price: 1 ml / €399.00

  Acris Antibodies GmbH

ERBB3 / HER3 antibody

AM09108PU c-erbB3 antibody staining of Human Small Intestine. Rabbit IgG SP71 Purified Hu P
0.1 ml / €290.00
  Acris Antibodies GmbH

ERBB3 / HER3 antibody

Human colon carcinoma stained with anti-c-erbB3 antibody Rabbit Purified Hu P
1 ml / €520.00
  Acris Antibodies GmbH

ERBB3 / HER3 antibody

Human colon carcinoma stained with anti-c-erbB3 antibody Rabbit Purified Hu P

Regular Price: 0.2 ml / €200.00

Special Price: 0.2 ml / €140.00

  Acris Antibodies GmbH

ERBB3 / HER3 (C-term) (incl. pos. control) antibody

Detection of Endogenous ErbB3: Whole cell lysates of EGF stimulated serum starved tumor cells (20.000 cells per lane) were applied to SDS-PAGE and transferred to PVDF membranes. Immunoblots were probed with mab erbB3-11A4 (0.5 µg/ ml) for 1h at RT and developed by ECL (exp. time: 30 sec). 
Lane 1: MDA-MB 231
Lane 2: MDA-MB 468
Lane 3: MCF-7
Lane 4: T47D
Lane 5: SW480
Lane 6: SW620 Mouse IgG1 11A4 Purified Hu WB
0.1 mg / €430.00
  Acris Antibodies GmbH

ERBB3 / HER3 (Cytopl. Dom.) (incl. pos. control) antibody

Detection of endogenous erbB3: Whole cell lysates of EGF stimulated serum starved tumor cells (20.000 cells per lane) were applied to SDS-PAGE and transferred to a PVDF membrane. The immunoblot was probed with mab erbB3-5A12 (0.5 µg/ ml) for 1h at RT and developed by ECL (exp. time: 30 sec). lane 1: MDA-MB 231; lane 2: MDA-MB 468; lane 3: MCF-7; lane 4: T47D; lane 5: SW480; lane 6: SW620 Mouse IgG1 5A12 Purified Hu ICC/IF, P, WB
0.1 mg / €430.00
  Acris Antibodies GmbH

ERBB3 / HER3 antibody

Western blot (WB) analysis of HER3 antibody (Cat.-No.: AP1222PU-N) in extracts from HeLa cells. Rabbit Aff - Purified Hu, Ms, Rt WB
0.1 mg / €350.00
  Acris Antibodies GmbH

ERBB3 / HER3 antibody

Figure 1. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Her3/ErbB3 Antibody Cat.-No. AP08087PU (Left) and the same antibody preincubated with blocking peptide (Right). Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, P, WB
0.1 ml / €270.00
  Acris Antibodies GmbH

ERBB3 / HER3 antibody

Figure 1. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Her3/ErbB3 Antibody Cat.-No. AP08087PU (Left) and the same antibody preincubated with blocking peptide (Right). Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, P, WB
50 µl / €230.00
  Acris Antibodies GmbH

ERBB3 / HER3 (1330-1342) antibody

Lung, respiratory epithelium: Formalin-Fixed Paraffin-Embedded (FFPE) Goat IgG Aff - Purified Bear, Bov, Bt, Eq, Hst, Hu, Mky, Ms, Por, Prim, Rt E, P, WB
50 µg / €470.00
  Acris Antibodies GmbH

ERBB3 / HER3 antibody

Immunofluorescence staining of methanol-fixed MCF cells using Her3/ErbB3 Antibody Rabbit Aff - Purified Hu ICC/IF
0.1 ml / €270.00
  Acris Antibodies GmbH

ERBB3 / HER3 antibody

Immunofluorescence staining of methanol-fixed MCF cells using Her3/ErbB3 Antibody Rabbit Aff - Purified Hu ICC/IF
50 µl / €230.00
  Acris Antibodies GmbH

ERBB3 / HER3 (N-term) antibody

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. Rabbit Ig Purified Hu, Ms P, WB

Regular Price: 0.4 ml / €370.00

Special Price: 0.4 ml / €231.00

  Acris Antibodies GmbH

ERBB3 / HER3 antibody

Formalin-fixed, paraffin-embedded human skeletal muscle stained with c-erbB3 Ab (Cat. SM1444P) using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of muscle cells. Mouse IgG1 SGP.1 Purified Hu C, E, F, IP

Regular Price: 0.1 mg / €540.00

Special Price: 0.1 mg / €350.00

  Acris Antibodies GmbH

ERBB3 / HER3 antibody

Western Blot: ErbB 3 Antibody [22590002] - Western Blot was performed using affinity purified SEQer ERBB3 antibody, aa(146-245) antibody. The lanes contain 5-30ug of a whole cell extract. Final concentration of antibodies = 0.1ug/ml (1:10,000 dilution). The blot was probed overnight with the SEQer ERBB3 antibody, aa(146-245) antibody. Blot was then washed according to protocol and probed with goat-anti-Rabbit-HRP conjugate at 1:5000 dilution, washed and developed using chemiluminescence (film exposure 5-30sec). The protein was detected as represented by the band shown. The detected protein band is estimated at 110 KDa, and the predicated molecular weight of ERBB3 is 140 KDa. The difference of molecular weight between empirical and predicted results are unclear. It could cause by post-translational modification, isotype, proteolytic degradation, gel artifacts or the other reasons. Rabbit Hu E, WB
50 µg / €440.00
  Novus Biologicals Inc.

ERBB3 / HER3 antibody

NB 100-2416 (0.1ug/ml) staining of Jurkat lysate (35ug protein in RIPA buffer) with (B) and without (A) blocking with the immunising peptide.  Primary incubation was 1 hour.  Detected by chemiluminescence. Goat IgG Aff - Purified Hu E, WB
0.1 mg / €400.00
  Novus Biologicals Inc.

ERBB3 / HER3 antibody

ERBB3 / HER3 Mouse IgG1 RTJ2 Purified Hu E, IP, P, WB
0.1 mg / €340.00
  Novus Biologicals Inc.

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