Immunofluorescent analysis of c-Met was performed on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0. 25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ c-Met recombinant rabbit monoclonal antibody (Product # 700261) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 goat anti-rabbit IgG secondary antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 phalloidin (Product # A12381) and Panel d is a merged image showing punctated staining on the membrane and panel e is a control without primary antibody. The images were captured using Nikon enabled with cool snap at 20X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A peptide corresponding to amino acids 1379-1390 of P08581.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1-5 ug/ml|
|Flow Cytometry (Flow)||1:100-1:500|
|Western Blot (WB)||1:500-1:5000|
* 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.
This antibody is predicted to react with primate based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
c-Met is tyrosine kinase receptor composed of a disulfide-linked heterodimer made of 45 kDa alpha-and 145 kDa beta-subunits found in many tissues. It expresses a transmembrane receptor-like protein, and is involved in regulating cellular proliferation, motility, morphogenesis, and apoptosis. c-Met encodes a high-affinity receptor for hepatocyte growth factor. Additionally, c-Met overexpression has been found in a wide variety of cancer types, particularly lung cancer.
IP-MS enrichment of MET (LFQ intensity): MET was enriched 168-fold from BT549 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and c-MET antibody (Part No. 700261). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
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