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|Antigen||Kinase insert Domain Receptor (A Type III Receptor tyrosine Kinase) (KDR) Antibodies|
|Epitope||C-Term, AA 1300-1367 Alternatives|
|Reactivity||Human, Mouse (Murine) Alternatives|
|Conjugate||This VEGFR2 antibody is un-conjugated Alternatives|
Immunocytochemistry (ICC), Immunofluorescence (IF), Western Blotting (WB)
|1 reference available|
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Product Details anti-VEGFR2 AntibodyTarget Details VEGFR2 Application Details Handling ProductDetails: References for anti-VEGFR2 antibody (ABIN151962) Images
|Purification||Immunogen affinity purified|
|Immunogen||A synthetic peptide made to a C-terminal region of the mouse VEGF Receptor 2 protein (between residues 1300-1367). [Swiss-Prot# P35918]|
Target Details VEGFR2Product Details anti-VEGFR2 Antibody Application Details Handling ProductDetails: References for anti-VEGFR2 antibody (ABIN151962) Images back to top
|Alternative Name||CD309 / VEGFR-2 / Flk-1 (KDR Antibody Abstract)|
|Background||Gene Symbol: KDR|
|Molecular Weight||Theoretical MW: 150 kDa|
|Pathways||RTK Signaling, Glycosaminoglycan Metabolic Process, Signaling Events mediated by VEGFR1 and VEGFR2, Growth Factor Binding, Regulation of long-term Neuronal Synaptic Plasticity, VEGF Signaling|
Application DetailsProduct Details anti-VEGFR2 Antibody Target Details VEGFR2 Handling ProductDetails: References for anti-VEGFR2 antibody (ABIN151962) Images back to top
|Application Notes||Western Blot 1:250-1:1000, Immunocytochemistry/Immunofluorescence 1:50-1:500This 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). The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.|
The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.
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 ProductDetails: References for anti-VEGFR2 antibody (ABIN151962) Images back to top
Tris-Glycine and 0.15M NaCl
Buffer contains: 0.05 % Sodium Azide
|Precaution of Use||This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.|
|Handling Advice||Do not freeze.|
|Storage Comment||Store at 4°C. Do not freeze.|
ProductDetails: References for anti-VEGFR2 antibody (ABIN151962)Product Details anti-VEGFR2 Antibody Target Details VEGFR2 Application Details Handling Images back to top
|Product cited in:||
Motiani, Savolainen, Eskelinen, Toivanen, Ishizu, Yli-Karjanmaa, Virtanen, Parkkola, Kapanen, Grönroos, Haaparanta-Solin, Solin, Savisto, Ahotupa, Löyttyniemi, Knuuti, Nuutila, Kalliokoski et al.: "Two weeks of moderate-intensity continuous training, but not high-intensity interval training, increases insulin-stimulated intestinal glucose uptake. ..." in: Journal of applied physiology (Bethesda, Md. : 1985), Vol. 122, Issue 5, pp. 1188-1197, 2017 Method employed by authors: Western Blotting (WB) (Sample species: Rat (Rattus)).
ImagesProduct Details anti-VEGFR2 Antibody Target Details VEGFR2 Application Details Handling ProductDetails: References for anti-VEGFR2 antibody (ABIN151962) back to top