This VEGF antibody is useful for Immunohistochemistry (frozen and paraffin-embedded sections), Western blot and Immunocytochemistry/Immunofluorescence. ELISA was reported in scientific literature. In IHC a dilution of 1:20-1:50 was used in an ABC method. However, depending on the staining conditions employed, we suggest that the final dilution should be determined by the user. We suggest an incubation period of 30-60 minutes at room temperature. High temperature treatment of formalin-fixed tissue sections using 1mM EDTA, pH 8.0 must be performed prior to the immunostaining.
VEGF (vascular endothelial growth factor) is a homodimeric, disulfide-linked glycoprotein growth factor that plays a critical role in angiogenesis, vasculogenesis and endothelial cell growth through induction of endothelial cell proliferation and blood vessels permeabilization, cell migration promotion as well as inhibition of apoptosis. VEGF can bind to FLT1/VEGFR1 and KDR/VEGFR2 receptors, heparan sulfate and heparin. Its isoforms VEGF189, VEGF165 and VEGF121 are widely expressed, whereas, other isoforms VEGF206 and VEGF145 are not very common. The basic isoform VEGF189 is cell-associated after secretion and is bound avidly by heparin and the extracellular matrix, although it may be released as a soluble form by heparin, heparinase or plasmin. VEGF bind to three tyrosine-kinase receptors, VEGFR-1, VEGFR-2 and VEGFR-3 which are expressed almost exclusively in endothelial cells. VEGFR-2 is the main angiogenic signal transducer for VEGF, while VEGFR-3 is specific for VEGF-C/-D (may gain VEGFR-2 binding ability via proteolytic processing) and is essential for lymphangiogenic signaling. VEGF is regulated by growth factors, cytokines, gonadotropins, nitric oxide, hypoxia, hypoglycemia and oncogenic mutations. Defects in VEGFA are linked to MVCD1 (microvascular complications of diabetes type 1) and VEGF polymorphisms are associated with susceptibility to multiple cancers, e.g., glioma, HCC, ovarian, bladder, prostate, breast cancer etc.
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FAQs for VEGF (8)
Why is the molecular weight of VEGF different from the similar antibody, for some companies the the molecular weight is 40KD)?
I can't comment on another company's antibody because I don't have any information about their products. I can tell you that VEGF is expressed in a variety of isoforms and is subject to various post-translational modifications that influence its apparent molecular weight in an SDS-PAGE gel compared to the theoretical molecular weight.
I would be very grateful if you could let me know a recommended dilution for the VEGF antibody antibody NB100-664
The recommended dilution for IHC is 1:20-1:50 in an ABC method. If instead you are testing on cell culture, I would recommend a starting optimization point of 1:10 -1:2000.
I would like to know the aproximate dilution of the VEGF Antibody (NB100-664) to use it in a Western blot assay.
In regards to your inquiry about our VEGF antibody (NB100-664), I am showing from the lab data that they have tested dilutions in the range of 1:500-1:1000 for this antibody. The optimal dilution should depend on your samples and the level of expression of VEGF, but a good starting point would be 1:1000 and then optimize from there.
What is the minimum amount of protein necessary for the detection of VEGF by Western blot using VEGF Antibody (NB100-664)?
I would suggest at minimum 10ug of protein per well but if you have enough sample you might want to run multiple wells with increasing amounts of protein to optimize the conditions in your hands. You could start with 5ug and do a couple of wells with 10ug, 20ug and 40ug. Also try using different dilutions of primary, start with 1:500-1:1000 and work your way up or down depending on what your blot looks like.
I am interested in buying an antibody to VEGF to detect VEGF in human umbilical vein endothelial cells for western blotting and was wondering if you might have any recommendations? I was thinking about NB100-664. Do you have a recommended amount to use for western blotting? Could I ask you what concentration the antibody is at since it says 0.1mg so what total volume do you send? Also is this the most commonly purchase one for western blotting do you know?
We do not have a recommended dilution noted for this specific antibody, but our general recommendation for Western blot is 1:1000-1:2000.This is a good starting point, but we always recommend optimization by the end user, as the optimal dilution is to some extent dependent on the particular sample/assay. The concentration for this product is stated as 1 mg/ml, so 0.1 mg would be supplied in 0.1 ml.
I am looking for a VEGF-A antibody for the IHC staining of mouse colon. Do you have any publications or application references for this product?
NB100-664 is one of our best sellers with great customer feedback and citations in at least 18 peer reviewed publications from highly esteemed researchers. As far as mouse tissues are concerned, I would recommend that you go through the following paper: Milanesi A, et al. Beta-Cell regeneration mediated by human bone marrow mesenchymal stem cells. PLoS One. 2012;7(8):e42177. doi: 10.1371/journal.pone.0042177. Epub 2012 Aug 7. PMID:22879915.
In the immunohistochemistry paraffin section protocol....the PBS buffer....should it be at certain pH?
For IHC application, different labs cites the pH of PBS buffer in the range of 7.1 - 7.6 but most commonly used pH is pH 7.4 (this is what we use in our lab). You may use this PBS (pH 7.4) for making permeablization buffer, antibody diluent buffers and for making wash buffer. For more on the protocol that we use in our lab and for IHC-P troubleshooting suggestions, you may visit: IHC-P Protocol and IHC-P Troubleshooting.
I was wondering what does Format 7C mean as far as anitbodies go?
7C is a fluorophore - it is useful for FLOW. The details of the fluor are (A=425, E=500)
VEGF: Vascular endothelial growth factor VEGF is homodimeric, disulfide-linked glycoprotein cytokine that serves as the ligand for FLT1 (VEGFR-1 receptor) and FLK1 (VEGFR-2 receptor) tyrosine kinases. It is a key modulator of physiological angiogenesis, vasculogenesis, and endothelial cell g... Read full blog post.
LYVE1 - It's Alive! It's a LYVE LYVE1 (lymphatic vessel endothelial hyaluronan receptor-1) is one of the most specific and widely used mammalian lymphatic endothelial markers.It is found in lymph nodes and at the luminal/abluminal surfaces of lymphatic vessels. It is a single-pass t... Read full blog post.
LOX: A prime enzyme LOX is a copper-dependent amine oxidase enzyme that executes post-translational oxidative deamination on peptidyl lysine residues in precursors of fibrous collagen and elastin. LOX is secreted into the extracellular environment in an inactive form, wh... Read full blog post.
VEGF Receptors, Angiogenesis and Cancer Vascular endothelial growth factor receptors 1 and 2 (VEGFR1 and VEGFR2) are related family members of the vascular endothelial growth factor (VEGF) family of membrane receptor tyrosine kinases. They are key regulators of physiological angiogenesis du... Read full blog post.
Controlling the HIF-1 Switch Hypoxia-inducible factor-1 is a major transcription factor composed of two subunits: HIF-1alpha and HIF-1 beta. Under normoxic conditions, HIF-1 alpha is targeted to proteosomal degradation via ubiquitination. On the other hand during hypoxic conditio... Read full blog post.