VEGF Antibody - BSA Free

Novus Biologicals | Catalog # NB100-2381

Novus Biologicals
100% Guarantee

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Bovine, Canine, Chicken, Equine, Guinea Pig

Cited:

Human, Mouse, Rat, Avian - Chicken

Applications

Validated:

Western Blot, ELISA

Cited:

Western Blot, ELISA

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all VEGF Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an internal region of the human VEGFA protein sequence (between residues 180-232). [Swiss-Prot: P15692]

Localization

Secreted

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for VEGF Antibody - BSA Free

Western Blot: VEGF AntibodyBSA Free [NB100-2381]

Western Blot: VEGF AntibodyBSA Free [NB100-2381]

Western Blot: VEGF Antibody [NB100-2381] - Detection of VEGFA doublet in CSF-IR/VEGFA chimera transfected lysate using.
VEGF Antibody - BSA Free

Western Blot: VEGF Antibody - BSA Free [NB100-2381] -

HIF-1 alpha protein and HIF-1 alpha target genes are elevated in SCs following retrovirus administration. Expression of mRNA for VP16, HIF-1 alpha, and beta -actin in transfected SCs was confirmed by PCR in VP16 and VP16-HIF SCs (A). Protein expression of VP16, HIF-1 alpha, HIF-2 alpha, VEGF, enolase, and protein loading controls (GFP or beta -actin) in transfected SCs was assessed by western blotting (B). Protein expression was normalized to the loading control. Relative intensity of VP16 expression is shown in C, and the fold change in protein expression relative to control SCs is shown for HIF-1 alpha (D) HIF-2 alpha (E), VEGF (F), and enolase (G). Values are as follows: VP16 (control: 0.0 +/- 0.0; VP16: 0.40 +/- 0.01; VP16-HIF: 0.41 +/- 0.00); HIF-1 alpha (control: 1.0 +/- 0.04; VP16: 1.0 +/- 0.04; VP16-HIF: 5.9 +/- 0.01); HIF-2 alpha (control: 1.0 +/- 0.01; VP16: 1.0 +/- 0.01; VP16-HIF: 1.0 +/- 0.02); VEGF (control: 1.0 +/- 0.01; VP16: 1.02 +/- 0.02; VP16-HIF: 2.2 +/- 0.01); enolase (control: 1.0 +/- 0.01; VP16: 1.0 +/- 0.04; VP16-HIF: 1.7 +/- 0.00); n = 3/group. Mean +/- SEM, *p < 0.0001. Figure Contributions: Veena Kandaswamy generated the VP16-HIF cells. Veena Kandaswamy performed the PCRs. Ying Dai performed the western blottings. Ying Dai and Caitlin Hill analyzed the data. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31488552), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for VEGF Antibody - BSA Free

Application
Recommended Usage

ELISA

1:500

Western Blot

1 ug/mL
Application Notes
In Western blot, a band is seen at ~42 kDa with a chimera transfected lysate representing the homodimer VEGFA. Another faint band is seen at ~21 kDa, representing the monomer form of VEGFA. 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. This antibody did not work in IHC.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.09% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: VEGF

Vascular endothelial growth factor (VEGF), also called VEGF-A and vascular permeability factor (VPF), is a secreted homodimeric glycoprotein belonging to the VEGF family with a role in stimulating angiogenesis and vasculogenesis (1,2). More specifically, VEGF-A secretion from most cell types contributes to promoting endothelial cell proliferation and migration, inhibiting apoptosis, increasing vascular permeability, and wound healing (1). The VEGF family consists of several members including VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, VEGF-F, and placenta growth factor (PLGF) (1-4). As a result of alternative splicing of the eight exon VEGFA gene, there are several VEGF-A protein isoforms of 121, 145, 165, 183, 189, and 206 amino acids (aa) in length, with VEGF121 and VEGF165 being the two most expressed isoforms (1,5). Full length VEGF-A monomer has a 26 aa signal sequence plus a 206 aa (VEGF206) sequence, with a theoretic molecular weight (MW) of 27 kDa, containing VEGF receptor 1 (VEGFR1) and VEGR2 binding sites and heparin-binding domains (1-3,5,6). VEGF121 lacks heparin affinity and binds the receptor tyrosine kinases (RTKs) VEGFR1 and VEGFR2, whereas VEGF165 has moderate affinity for heparin and, in addition to being a ligand for VEGFR1 and VEGFR2, can also bind the co-receptors neuropilin 1 (NRP1) and NRP2 (1,5). Hypoxia and hypoxia-related genes such as HIF-1, EGF, and PDGF are major regulators angiogenesis and VEGF expression (1,3). VEGF signaling initiated by ligand binding to its receptors results in activation of different pathways including PI3K and MAPK and ultimately guides endothelial cell proliferation, migration, and survival (1,3). While VEGF plays an important role in promoting normal angiogenesis and blood vessel formation, its expression is often upregulated in tumors and other angiogenesis-related pathologies like osteroarthritis (OA) (1-5,7). Given its function, VEGF and its receptors have become a therapeutic target for treating cancer and blocking angiogenesis (4,5,7). A recombinant humanized monoclonal anti-VEGFA antibody called bevacizumab (Avastin) was first approved by the FDA in 2004 for the treatment of a number of cancers (1-3,5). Cancer patients may experience resistance to anti-VEGF antibodies and, as such, clinical studies are exploring combination treatment options with chemotherapies and immune-checkpoint inhibitors (3,5).

References

1. Melincovici CS, Bosca AB, susman S, et al. Vascular endothelial growth factor (VEGF) - key factor in normal and pathological angiogenesis. Rom J Morphol Embryol. 2018;59(2):455-467.

2. Shaik F, Cuthbert GA, Homer-Vanniasinkam S, Muench SP, Ponnambalam S, Harrison MA. Structural Basis for Vascular Endothelial Growth Factor Receptor Activation and Implications for Disease Therapy. Biomolecules. 2020;10(12):1673. https://doi.org/10.3390/biom10121673

3. Apte RS, Chen DS, Ferrara N. VEGF in Signaling and Disease: Beyond Discovery and Development. Cell. 2019;176(6):1248-1264. https://doi.org/10.1016/j.cell.2019.01.021

4. Matsumoto K, Ema M. Roles of VEGF-A signalling in development, regeneration, and tumours. J Biochem. 2014;156(1):1-10. https://doi.org/10.1093/jb/mvu031

5. Itatani Y, Kawada K, Yamamoto T, Sakai Y. Resistance to Anti-Angiogenic Therapy in Cancer-Alterations to Anti-VEGF Pathway. Int J Mol Sci. 2018;19(4):1232. Published 2018 Apr 18. doi:10.3390/ijms19041232

6. Uniprot (P15692)

7. Hamilton JL, Nagao M, Levine BR, Chen D, Olsen BR, Im HJ. Targeting VEGF and Its Receptors for the Treatment of Osteoarthritis and Associated Pain. J Bone Miner Res. 2016;31(5):911-924. https://doi.org/10.1002/jbmr.2828

Long Name

Vascular Endothelial Growth Factor

Alternate Names

MVCD1, VAS, Vasculotropin, VEGF-A, VEGFA, VPF

Gene Symbol

VEGFA

Additional VEGF Products

Product Documents for VEGF Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for VEGF Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for VEGF Antibody - BSA Free

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Protocols

View specific protocols for VEGF Antibody - BSA Free (NB100-2381):

VEGF Antibody:
Western Blot Protocol

1. Perform SDS-PAGE (4-12%) on samples to be analyzed.

2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.

3. Rinse membrane with dH2O and then 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.

4. Rinse the blot in TBS for approximately 5 minutes.

5. Block the membrane using 5% non-fat dry milk + 1% BSA in TBS for 1 hour at room temperature.

6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.

7. Dilute the rabbit anti-VEGFA primary antibody (NB 100-2381) in blocking buffer and incubate 1 hour at room temperature.

8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.

9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) and incubate 1 hour at room temperature.

10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).

11. Apply the detection reagent of choice in accordance with the manufacturer's instructions (we used BioFX Super Plus ECL).

Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for VEGF Antibody - BSA Free

Showing  1 - 4 of 4 FAQs Showing All

  • Q: I am looking for an antibody for detection of VEGFA in bovine samples in an ELISA-type assay. I found NB100-2381H. Do you know if this product has been specifically tested with bovine or if cross-reactivity is merely predicted based on homology? Also, do you know if ELISA in this case refers to detection of actual endogenous protein in a sample or if the assay was peptide-based?

    A: We verified ELISA as an application based on a customer review. They used the antibody to identify the recombinant protein. It has not yet been tested to detect the endogenous protein in other samples like blood or plasma. Bovine is listed as reactive species based on homology, but we will fully guarantee detection of the bovine protein with this product. We will also guarantee the ELISA application. This will go for both the HRP conjugated antibody NB100-2381H and the unconjugated antibody NB100-2381.

  • Q: We are looking for a VEGFA antibody for use with porcine. Do you know if this antibody has actually been tested with porcine samples, or if cross-reactivity is just predicted based on homology? Is there any info or data regarding use with porcine?

    A: The chimera transfected lysate shows on our website that is actually a porcine aorta endothelial cell line, which has been transfected with human VEGFR2. The antibody is detecting both the human protein and the endogenous porcine protein. We have received feedback from customers that NB100-2381 works in porcine tissues.

  • Q: What is the heparin binding activity of FGF and VEGF?

    A: These proteins are not assayed for their ability to bind Heparin. More information about the FGF family of growth factors is available in this review article: Basilico, C. (1992) Adv. Can. Res. 59:115.

  • Q: Why is the molecular weight of VEGF different from the similar antibody, for some companies the the molecular weight is 40KD)?

    A: 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.

  • Q: I am looking for an antibody for detection of VEGFA in bovine samples in an ELISA-type assay. I found NB100-2381H. Do you know if this product has been specifically tested with bovine or if cross-reactivity is merely predicted based on homology? Also, do you know if ELISA in this case refers to detection of actual endogenous protein in a sample or if the assay was peptide-based?

    A: We verified ELISA as an application based on a customer review. They used the antibody to identify the recombinant protein. It has not yet been tested to detect the endogenous protein in other samples like blood or plasma. Bovine is listed as reactive species based on homology, but we will fully guarantee detection of the bovine protein with this product. We will also guarantee the ELISA application. This will go for both the HRP conjugated antibody NB100-2381H and the unconjugated antibody NB100-2381.

  • Q: We are looking for a VEGFA antibody for use with porcine. Do you know if this antibody has actually been tested with porcine samples, or if cross-reactivity is just predicted based on homology? Is there any info or data regarding use with porcine?

    A: The chimera transfected lysate shows on our website that is actually a porcine aorta endothelial cell line, which has been transfected with human VEGFR2. The antibody is detecting both the human protein and the endogenous porcine protein. We have received feedback from customers that NB100-2381 works in porcine tissues.

  • Q: What is the heparin binding activity of FGF and VEGF?

    A: These proteins are not assayed for their ability to bind Heparin. More information about the FGF family of growth factors is available in this review article: Basilico, C. (1992) Adv. Can. Res. 59:115.

  • Q: Why is the molecular weight of VEGF different from the similar antibody, for some companies the the molecular weight is 40KD)?

    A: 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.

  • Q: I am looking for an antibody for detection of VEGFA in bovine samples in an ELISA-type assay. I found NB100-2381H. Do you know if this product has been specifically tested with bovine or if cross-reactivity is merely predicted based on homology? Also, do you know if ELISA in this case refers to detection of actual endogenous protein in a sample or if the assay was peptide-based?

    A: We verified ELISA as an application based on a customer review. They used the antibody to identify the recombinant protein. It has not yet been tested to detect the endogenous protein in other samples like blood or plasma. Bovine is listed as reactive species based on homology, but we will fully guarantee detection of the bovine protein with this product. We will also guarantee the ELISA application. This will go for both the HRP conjugated antibody NB100-2381H and the unconjugated antibody NB100-2381.

  • Q: We are looking for a VEGFA antibody for use with porcine. Do you know if this antibody has actually been tested with porcine samples, or if cross-reactivity is just predicted based on homology? Is there any info or data regarding use with porcine?

    A: The chimera transfected lysate shows on our website that is actually a porcine aorta endothelial cell line, which has been transfected with human VEGFR2. The antibody is detecting both the human protein and the endogenous porcine protein. We have received feedback from customers that NB100-2381 works in porcine tissues.

  • Q: What is the heparin binding activity of FGF and VEGF?

    A: These proteins are not assayed for their ability to bind Heparin. More information about the FGF family of growth factors is available in this review article: Basilico, C. (1992) Adv. Can. Res. 59:115.

  • Q: Why is the molecular weight of VEGF different from the similar antibody, for some companies the the molecular weight is 40KD)?

    A: 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.

  • Q: I am looking for an antibody for detection of VEGFA in bovine samples in an ELISA-type assay. I found NB100-2381H. Do you know if this product has been specifically tested with bovine or if cross-reactivity is merely predicted based on homology? Also, do you know if ELISA in this case refers to detection of actual endogenous protein in a sample or if the assay was peptide-based?

    A: We verified ELISA as an application based on a customer review. They used the antibody to identify the recombinant protein. It has not yet been tested to detect the endogenous protein in other samples like blood or plasma. Bovine is listed as reactive species based on homology, but we will fully guarantee detection of the bovine protein with this product. We will also guarantee the ELISA application. This will go for both the HRP conjugated antibody NB100-2381H and the unconjugated antibody NB100-2381.

  • Q: We are looking for a VEGFA antibody for use with porcine. Do you know if this antibody has actually been tested with porcine samples, or if cross-reactivity is just predicted based on homology? Is there any info or data regarding use with porcine?

    A: The chimera transfected lysate shows on our website that is actually a porcine aorta endothelial cell line, which has been transfected with human VEGFR2. The antibody is detecting both the human protein and the endogenous porcine protein. We have received feedback from customers that NB100-2381 works in porcine tissues.

  • Q: What is the heparin binding activity of FGF and VEGF?

    A: These proteins are not assayed for their ability to bind Heparin. More information about the FGF family of growth factors is available in this review article: Basilico, C. (1992) Adv. Can. Res. 59:115.

  • Q: Why is the molecular weight of VEGF different from the similar antibody, for some companies the the molecular weight is 40KD)?

    A: 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.

Showing  1 - 4 of 4 FAQs Showing All
View all FAQs for Antibodies