Application Notes | Western Blot 1:100-1:2000, Immunohistochemistry, Immunocytochemistry/Immunofluorescence 1:500, Immunohistochemistry-ParaffinThis VEGF Receptor 1 antibody is useful for Western blot, where a band is seen at ~180-230 kDa (the theoretical molecular weight of mouse VEGFR1 is ~150 kDa and human VEGFR1 is ~151 kDa). The difference in MWs is likely due to glycosylation or other post translational modifications. With CSF-1R/VEGFR2 chimera transfected lysates a doublet is seen at ~150 kDa representing VEGFR-2. An unknown band is also seen at ~50 kDa. Optimal working dilutions should be determined by the investigator. |
Comment |
The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo. |
Protocol |
Protocol specific for VEGF Receptor 1 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-VEGFR1 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. |
Restrictions | For Research Use only |
Format | Liquid |
Concentration | 1 mg/mL |
Buffer |
Tris-Glycine and 0.15M NaCl Buffer contains: 0.05 % Sodium Azide |
Preservative | 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 | 4 °C |
Storage Comment | Store at 4°C. Do not freeze. |
Supplier Images |
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Product cited in: |
Puglisi, Puppin, Pegolo, Andreetta, Pascoletti, DAurizio, Pandolfi, Fasola, Piga, Damante, Di Loreto: "Expression of periostin in human breast cancer." in: Journal of clinical pathology, Vol. 61, Issue 4, pp. 494-8, 2008 (PubMed).
Ptaszynska, Pendrak, Bandle, Stracke, Roberts: "Positive feedback between vascular endothelial growth factor-A and autotaxin in ovarian cancer cells." in: Molecular cancer research : MCR, Vol. 6, Issue 3, pp. 352-63, 2008 (PubMed). (Sample species: Human). Further details: Western Blotting Takahashi, Hattori, Iwamatsu, Takizawa, Shibuya: "A novel snake venom vascular endothelial growth factor (VEGF) predominantly induces vascular permeability through preferential signaling via VEGF receptor-1." in: The Journal of biological chemistry, Vol. 279, Issue 44, pp. 46304-14, 2004 (PubMed). Rahimi, Dayanir, Lashkari: "Receptor chimeras indicate that the vascular endothelial growth factor receptor-1 (VEGFR-1) modulates mitogenic activity of VEGFR-2 in endothelial cells." in: The Journal of biological chemistry, Vol. 275, Issue 22, pp. 16986-92, 2000 (PubMed). |
Background publications |
Gluzman-Poltorak, Cohen, Shibuya, Neufeld: "Vascular endothelial growth factor receptor-1 and neuropilin-2 form complexes." in: The Journal of biological chemistry, Vol. 276, Issue 22, pp. 18688-94, 2001 (PubMed).
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