Immunofluorescence analysis of ErbB3 was done on 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ErbB3 (RTJ2) Mouse Monoclonal Antibody (MA1860) at 2ug/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Synthetic Peptide: E(1004) L E P E L D L D L D L E(1016)|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Flow Cytometry (Flow)||0.5-2 ug per 10^6 cells|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (IHC)||6 - 8 µg/mL|
|Immunohistochemistry (Paraffin) (IHC (P))||5 µg/ml|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-860 detects Her-3 from human and mouse samples.
MA1-860 has been successfully used in FACS, immunoprecipitation, immunohistochemical, immunofluorescence and ELISA procedures. By immunoprecipitation, this antibody detects ~185 kDa protein, representing Her-3 from human tissue extracts.This antibody has also been used in immunohistochemical, immunofluorescence and ELISA procedures using human tissues.
MA1-860 immunizing peptide corresponds to amino acid residues 1004 to 1016 of human cytoplasmic domain of HER-3 protein.
Her-3, also known as c-erbB-3, is a Type 1 membrane protein found in numerous cell types. It is a member of the EGF receptor protein family, and is activated by neuregulins and NTAK. Her-3 levels have been found to be elevated in certain human tumor cell lines suggesting that it may play a role in some malignancies. Her-3 is a receptor for heregulin and is capable of mediating HGL-stimulated tyrosine phosphorylation of itself.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Heregulin and retinoids synergistically induce branching morphogenesis of breast cancer cells cultivated in 3D collagen gels.
MA1-860 was used in immunocytochemistry to assess the morphogenetic activities of c-erbB and retinoids on human mammary epithelial cells
|Offterdinger M,Schneider SM,Grunt TW||Journal of cellular physiology (195:260)||2003|
Neuregulin expression, function, and signaling in human ovarian cancer cells.
MA1-860 was used in western blot to investigate the characteristics of neuregulin in human ovarian cancer cells
|Gilmour LM,Macleod KG,McCaig A,Sewell JM,Gullick WJ,Smyth JF,Langdon SP||Clinical cancer research : an official journal of the American Association for Cancer Research (8:3933)||2002|