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|Application / Reactivity||Rat (Rattus)||Mouse (Murine)|
|ELISA||36 Antibodies||69 Antibodies|
|Flow Cytometry (FACS)||2 Antibodies||40 Antibodies|
|Immunochromatography (IC)||2 Antibodies||3 Antibodies|
|Immunocytochemistry (ICC)||9 Antibodies|
|Immunofluorescence (IF)||44 Antibodies|
|Immunofluorescence (Paraffin-embedded Sections) (IF (p))||22 Antibodies|
|Immunohistochemistry (IHC)||67 Antibodies|
|Immunohistochemistry (Frozen Sections) (IHC (fro))||3 Antibodies|
|Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))||31 Antibodies|
|Western Blotting (WB)||85 Antibodies|
|Biochemical Assay (BCA)||1 Antibodies|
|Blocking Reagent (BR)||2 Antibodies|
|ELISA (Detection)||6 Antibodies|
|Enzyme Immunoassay (EIA)||1 Antibodies|
|Antigen||Kinase insert Domain Receptor (A Type III Receptor tyrosine Kinase) (KDR) Antibodies|
|Epitope||C-Term, AA 1300-1367 Alternatives|
|Reactivity||Human, Mouse (Murine), Rat (Rattus) Alternatives|
|Conjugate||This VEGFR2 antibody is un-conjugated Alternatives|
Immunocytochemistry (ICC), Immunofluorescence (IF), Western Blotting (WB)
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Product Details anti-VEGFR2 AntibodyTarget Details VEGFR2 Application Details Handling Images
|Cross-Reactivity (Details)||Predicted to react with rat,based on 100% sequence homology.|
|Immunogen||A synthetic peptide made to a C-terminal region of the mouse VEGF Receptor 2 protein (between residues 1300-1367).|
Target Details VEGFR2Product Details anti-VEGFR2 Antibody Application Details Handling Images back to top
|Alternative Name||CD309 / VEGFR-2 / Flk-1 (KDR Antibody Abstract)|
VEGF receptor 2 is a member of a receptor tyrosine kinase family whose activationplays an essential role in a large number of biological processes such as embryonicdevelopment, wound healing, cell proliferation, migration and differentiation. Like othergrowth factor receptors, upon ligand binding VEGF receptor 2 dimerises and isautophosphorylated on multiple tyrosine residues. These sites can be involved in theregulation of kinase activity or serve as binding sites for SH2 and phosphotyrosinebinding containing signaling proteins. Phosphorylation of Tyrosines 1054 and 1059 inthe activation loop is required for activation of VEGF receptor 2 and its intrinsic tyrosinekinase activity. Alternate Names: anti-VEGFR2 antibody, anti-VEGFR 2 antibody, anti-Protein-tyrosine kinase receptorflk-1 antibody, anti-Fetal liver kinase 1 antibody, anti-Kinase NYK antibody, anti-FLK1antibody, anti-KDR antibody, anti-Kinase insert domain receptor antibody, anti-KRD1antibody, anti-Ly73 antibody, anti-Protein tyrosine kinase receptor FLK1 antibody,anti-Vascular endothelial growth factor receptor 2 antibody.
Gene Symbol: KDR
|Research Area||Cytokines, Receptors|
Application DetailsProduct Details anti-VEGFR2 Antibody Target Details VEGFR2 Handling Images back to top
This VEGF Receptor 2 antibody is useful for Immunocytochemistry/Immunofluorescence and Western blot, where a doublet is seen at ~150 kDa. In ICC/IF nuclear and membrane staining can be seen in V6.5 mouse embryonic stem cells (Catalog No. NBP1-41162).
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:50-1:500, Western Blot 1:250-1:1000
Western blot Protocol for VEGF Receptor 2 Antibody
Western Blot Protocol:
1. Perform SDS-PAGE (3-8 %) on samples to be analyzed, loading 50 µg of total protein per lane.
. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
. Stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
. Rinse the blot in TBS for approximately 5 minutes.
. Block the membrane using 5 % non-fat dry milk + 0.5 % BSA in TBS for 1 hour.
. Dilute the rabbit anti-VEGFR2 primary antibody in blocking buffer and incubate 2 hours at room temperature.
. Wash the membrane in water for 5 minutes and apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) and incubate 1 hour at room temperature.
. Wash the blot in TBS containing 0.05-0.1 % Tween-20 for 10-20 minutes.
. Wash the blot in type I water for an additional 10-20 minutes (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturer's instructions (Amersham's ECL is the standard reagent used).Note: Tween-20 can be added to the blocking buffer at a final concentration of 0.05-0.2 %, provided it does not interfere with antibody-antigen binding.Immunocytochemistry/Immunofluorescence Protocol for VEGF Receptor 2 Antibody Immunocytochemistry ProtocolCulture cells to appropriate density in 35 mm culture dishes or 6-well plates.
. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 30 minutes.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
|Restrictions||For Research Use only|
HandlingProduct Details anti-VEGFR2 Antibody Target Details VEGFR2 Application Details Images back to top
|Buffer||Tris-glycine, 150 mM NaCl, Sodium Azide|
|Precaution of Use||WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.|
|Handling Advice||Do not freeze.|
ImagesProduct Details anti-VEGFR2 Antibody Target Details VEGFR2 Application Details Handling back to top