Mouse VEGF-B 167/186 Antibody

R&D Systems | Catalog # AF590

R&D Systems

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Rat

Applications

Validated:

Western Blot, Blockade of Receptor-ligand Interaction

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all VEGF-B Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant mouse VEGF-B186
Pro22-Ala207
Accession # P49766

Specificity

Detects mouse VEGF-B

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Mouse VEGF-B 167/186 Antibody

Detection of VEGF-B 167/186 by Western Blot

Detection of VEGF-B 167/186 by Western Blot

ROS-triggered VEGF-B protein acts on the VEGFR-1 expressed on activated T cells.(A) VEGF-B protein levels in activated CD8+ T cells treated with N-acetyl-L-cysteine (NAC) (5 μM) or glutathione (GSH) (5 μM); n = 6. (B) VEGF-B protein levels in activated CD8+ T cells treated with H2O2 at a gradient concentration; n = 3. (C) Alkaline phosphatase–labeled VEGF-B protein (VEGF-B-AP) binding in NA or activated CD4+ or CD8+ T cells. Scale bar: 2 μm. (D and E) The phosphorylation levels of VEGFR-1 (phospho-tyrosine), PI3K (P85), or ERK1/2 in activated CD8+ T cells treated by anti–VEGF-B ( alpha VB) or anti–VEGFR-1 ( alpha VEGFR-1) antibodies; n = 3. Data are shown as mean ± SEM. P values were calculated using 2-tailed unpaired t test in A and 1-way ANOVA with Holm-Šídák’s post hoc test in B. *P < 0.05, ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39145452), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of VEGF-B 167/186 by Flow Cytometry

Detection of VEGF-B 167/186 by Flow Cytometry

Autocrine VEGF-B signaling is essential for the antitumor immunity of CD8+ T cells.(A–D) The tumor growth curve of MC38 cells or B16 cells on Vegfb WT or -cKO or Flt1 WT or -cKO mice by subcutaneous injection; n = 5. (E and F) Flow cytometry (FCM) analysis for the ratio of tumor-infiltrating CD8+ T cells in MC38 tumors on Vegfb WT or -cKO mice. (E) Cell numbers and (F) annexin V+ ratio of tumor-infiltrating CD8+ T cells were calculated; n = 5. (G and H) FCM analysis for the ratio of tumor-infiltrating CD8+ T cells in MC38 tumors on Flt1 WT or -cKO mice. (G) Cell numbers and (H) annexin V+ ratio of tumor-infiltrating CD8+ T cells were calculated; n = 5. (I) CD8+ Tn cells from Vegfb WT or -cKO OT-1 cells were activated, followed by Senp2 overexpression, and then these OT-1 T cells were transferred (i.v.) to recipient mice inoculated with MC38-OVA cells. (J) The tumor growth curves were recorded; n = 5. (K) The H-2Kb+ ratio in tumor-infiltrating CD8+ T cells and (L) the annexin V+ ratio of H-2Kb+ CD8+ T cells were calculated; n = 5. Data are shown as mean ± SEM. P values were calculated using 2-way ANOVA with Bonferroni’s post hoc test in A–D and J–L and 2-tailed unpaired t test in E–H. **P < 0.01, ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39145452), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of VEGF-B 167/186 by Western Blot

Detection of VEGF-B 167/186 by Western Blot

TCR activation induces VEGF-B protein expression in T cells.(A) Relative mRNA levels of the secreted factors in CD8+ Tn cells treated with or without (NA) alpha CD3/CD28 for 3 days; n = 3. Prf1, Ifng, Gzmb, and Tnf were positive indicators for T cell activation. (B) VEGF-B protein levels in NA or activated CD4+ and CD8+ T cells; n = 3. (C) Immunofluorescence images of VEGF-B in NA or activated CD4+ or CD8+ T cells. DAPI staining is in blue. Scale bar: 2 μm. (D) The mean fluorescent intensity (MFI) of VEGF-B in NA or activated CD4+ or CD8+ T cells; n = 3. (E) The population of VEGF-B+ and IFN-gamma + in NA or activated CD8+ T cells; n = 3. (F) VEGF-B protein levels in culture supernatants of NA or activated CD4+ or CD8+ T cells; n = 3. (G) The population of VEGF-B+ and H-2Kb+ cells in CD8+ T cells from the spleen of OT-1 mice infected by LM-OVA for 3 days; n = 5. (H) The population of VEGF-B+ cells in CD4+ or CD8+ T cells from colon tissue with DSS-induced colitis; n = 6. Data are shown as mean ± SEM. P values were calculated using 2-tailed unpaired t test in F. *P < 0.05, **P < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39145452), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of VEGF-B 167/186 by Flow Cytometry

Detection of VEGF-B 167/186 by Flow Cytometry

Autocrine VEGF-B signaling is essential for the antitumor immunity of CD8+ T cells.(A–D) The tumor growth curve of MC38 cells or B16 cells on Vegfb WT or -cKO or Flt1 WT or -cKO mice by subcutaneous injection; n = 5. (E and F) Flow cytometry (FCM) analysis for the ratio of tumor-infiltrating CD8+ T cells in MC38 tumors on Vegfb WT or -cKO mice. (E) Cell numbers and (F) annexin V+ ratio of tumor-infiltrating CD8+ T cells were calculated; n = 5. (G and H) FCM analysis for the ratio of tumor-infiltrating CD8+ T cells in MC38 tumors on Flt1 WT or -cKO mice. (G) Cell numbers and (H) annexin V+ ratio of tumor-infiltrating CD8+ T cells were calculated; n = 5. (I) CD8+ Tn cells from Vegfb WT or -cKO OT-1 cells were activated, followed by Senp2 overexpression, and then these OT-1 T cells were transferred (i.v.) to recipient mice inoculated with MC38-OVA cells. (J) The tumor growth curves were recorded; n = 5. (K) The H-2Kb+ ratio in tumor-infiltrating CD8+ T cells and (L) the annexin V+ ratio of H-2Kb+ CD8+ T cells were calculated; n = 5. Data are shown as mean ± SEM. P values were calculated using 2-way ANOVA with Bonferroni’s post hoc test in A–D and J–L and 2-tailed unpaired t test in E–H. **P < 0.01, ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39145452), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of VEGF-B 167/186 by Western Blot

Detection of VEGF-B 167/186 by Western Blot

ROS-triggered VEGF-B protein acts on the VEGFR-1 expressed on activated T cells.(A) VEGF-B protein levels in activated CD8+ T cells treated with N-acetyl-L-cysteine (NAC) (5 μM) or glutathione (GSH) (5 μM); n = 6. (B) VEGF-B protein levels in activated CD8+ T cells treated with H2O2 at a gradient concentration; n = 3. (C) Alkaline phosphatase–labeled VEGF-B protein (VEGF-B-AP) binding in NA or activated CD4+ or CD8+ T cells. Scale bar: 2 μm. (D and E) The phosphorylation levels of VEGFR-1 (phospho-tyrosine), PI3K (P85), or ERK1/2 in activated CD8+ T cells treated by anti–VEGF-B ( alpha VB) or anti–VEGFR-1 ( alpha VEGFR-1) antibodies; n = 3. Data are shown as mean ± SEM. P values were calculated using 2-tailed unpaired t test in A and 1-way ANOVA with Holm-Šídák’s post hoc test in B. *P < 0.05, ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39145452), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of VEGF-B 167/186 by Immunocytochemistry/ Immunofluorescence

Detection of VEGF-B 167/186 by Immunocytochemistry/ Immunofluorescence

TCR activation induces VEGF-B protein expression in T cells.(A) Relative mRNA levels of the secreted factors in CD8+ Tn cells treated with or without (NA) alpha CD3/CD28 for 3 days; n = 3. Prf1, Ifng, Gzmb, and Tnf were positive indicators for T cell activation. (B) VEGF-B protein levels in NA or activated CD4+ and CD8+ T cells; n = 3. (C) Immunofluorescence images of VEGF-B in NA or activated CD4+ or CD8+ T cells. DAPI staining is in blue. Scale bar: 2 μm. (D) The mean fluorescent intensity (MFI) of VEGF-B in NA or activated CD4+ or CD8+ T cells; n = 3. (E) The population of VEGF-B+ and IFN-gamma + in NA or activated CD8+ T cells; n = 3. (F) VEGF-B protein levels in culture supernatants of NA or activated CD4+ or CD8+ T cells; n = 3. (G) The population of VEGF-B+ and H-2Kb+ cells in CD8+ T cells from the spleen of OT-1 mice infected by LM-OVA for 3 days; n = 5. (H) The population of VEGF-B+ cells in CD4+ or CD8+ T cells from colon tissue with DSS-induced colitis; n = 6. Data are shown as mean ± SEM. P values were calculated using 2-tailed unpaired t test in F. *P < 0.05, **P < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39145452), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of VEGF-B 167/186 by Western Blot

Detection of VEGF-B 167/186 by Western Blot

Autocrine VEGF-B is required for the survival of activated T cells.(A–D) sh-Vegfb or negative control (sh-NC) CD8+ OT-1 Tn cells were transferred into recipients infected with LM-OVA. The H-2Kb+ ratio in CD8+ T cells from the blood (B) or spleen (C) of recipients; n = 5. (D) The LM CFUs in the spleen and liver of the infected recipients; n = 5. (E) VEGF-B and VEGFR-1 protein levels in WT and cKO T cells; n = 3. (F) The MFI of CD69, CD25, and CD44 in Vegfb or Flt1 WT and cKO CD8+ Tn cells with TCR activation; n = 3. (G) The annexin V+ and PI+ ratio in VEGF-B–treated (VB-treated) Vegfb or Flt1 WT and cKO CD8+ T cells; n = 3. (H) CD8+ Tn cells were activated to induce memory T cells (Tm) by IL-15. The numbers and ratios of Tm (CD44+CD62L+) or TE (CD44+CD62L–) cells with IgG or alpha VB treatment were calculated; n = 8. (I–M) Vegfb or Flt1 WT and cKO CD8+ OT-1 T cells were transferred to recipients, followed by LM-OVA infection. (J and K) H-2Kb+ ratio in CD8+ T cells from the blood of recipients; n = 5. (L and M) The memory precursor (KLRG1–CD127A+) ratio in H-2Kb+ CD8+ T cells from the spleen of recipients on day 7 of prime infection; n = 5. Data are shown as mean ± SEM. P values were calculated using 2-tailed unpaired t test in C, D, H, L, and M and 2-way ANOVA with Holm-Šídák’s post-hoc test in B, G, J, and K. *P < 0.05, **P < 0.01, ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39145452), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse VEGF-B 167/186 Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.04-0.1 µg/mL of this antibody will block 50% of the binding of 5 ng/mL of Recombinant Mouse VEGF-B186 (Catalog # 767-VE) to immobilized Recombinant Human VEGF R1/Flt-1 Fc Chimera (Catalog # 321-FL) coated at 4 µg/mL (100 µL/well). This antibody can also block the binding of Recombinant Mouse VEGF-B167 (Catalog # 2595-VE) to immobilized Neuropilin‑1 Fc Chimera.

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse VEGF-B167 (Catalog # 2595-VE)
Recombinant Mouse VEGF-B186 (Catalog # 767-VE)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: VEGF-B

Vascular endothelial growth factor B (VEGF-B; also known as VFR) is a member of the VEGF-PDGF supergene family of growth factor molecules (1 - 4). Five mouse members have been identified, including VEGF-A, -B, -C, -D, and PlGF(-2) (1, 5). VEGF family members are disulfide-linked homo- and heterodimeric proteins that are important regulators of vasculogenesis and lymphangiogenesis. Mouse VEGF-B has two isoforms, a 32 kDa single chain and a 21 kDa single chain form (6, 7). The long form (VEGF-B186) is 207 amino acids (aa) in length, with a 21 aa signal sequence and a 186 aa mature region. The short form (VEGF-B167) is 188 aa in length, with a 21 aa signal sequence and a 167 aa mature segment. Each mature isoform shows the same N-terminal 94 aa that contains a cysteine knot VEGF homology domain (6 - 8). VEGF-B186 is O-glycosylated; VEGF-B167 is not. VEGF-B167 binds heparin; VEGF-B186 does not. Thus, VEGF-B186 is secreted and freely diffusible in tissues (7). However, the VEGF-B167 isoform is the predominant form in tissue (9). Mouse VEGF-B186 is 93% and 87% aa identical to bovine and human VEGF-B186, respectively; mouse VEGF-B167 is 90% and 88% aa identical to bovine and human VEGF-B167, respectively. The mouse VEGF-B167 homodimer is 42 kDa in size, while the VEGF-B186 homodimer is 62 kDa in size. Unlike VEGF167, VEGF-B186 undergoes proteolytic processing that creates a partially processed 48 kDa homodimer and a fully processed 32 kDa homodimer. Processing appears to occur at Arg127 of the mature form (10). Both forms of VEGF-B can heterodimerize with VEGF (7). Both VEGF-B isoforms bind to VEGF receptor 1 (VEGF R1), but not VEGF R2 or VEGF R3 (11). VEGF-B167 also binds neuropilin-1, but only the 127 aa processed form of VEGF-B186 binds neuropilin-1 (10). As a dimer, full length VEGF-B186 does not interact with neuropilin-1, while any dimer that contains the processed VEGF-B127 subunit will interact with neuropilin-1 (10). The importance of differential neuropilin binding is unclear. VEGF-B deficient mice display an atrial conduction deficit (12). On endothelial cells, ligation of VEGF R1 by VEGF-B has been shown to regulate the expression and activity of urokinase type plasminogen activator and plasminogen activator inhibitor 1 (11).

References

  1. Li, X. and U. Eriksson (2001) Int. J. Biochem Cell Biol. 33:421.
  2. Olofsson, B. et al. (1999) Curr. Opin. Biotechnol. 10:528.
  3. Clauss, M. (2000) Semin. Thromb. Hemost. 26:561.
  4. Matsumoto, T. and L. Claesson-Welsh (2001) Sci STKE Dec. 11(112):RE21.
  5. DiPalma, T. et al. (1996) Mamm. Genome 7:6.
  6. Olofsson, B. et al. (1996) Proc. Natl. Acad. Sci. USA 93:2576.
  7. Olofsson, B. et al. (1996) J. Biol. Chem. 271:19310.
  8. Twonson, S. et al. (1996) Biochem. Biophys. Res. Commun. 220:922.
  9. Li, X. et al. (2001) Growth Factors 19:49.
  10. Makinen, T. et al. (1999) J. Biol. Chem. 274:21217.
  11. Olofsson, B. et al. (1998) Proc. Nat. Acad. Sci. USA 95:11709.
  12. Aase, K. et al. (2001) Circulation 104:358.

Long Name

Vascular Endothelial Growth Factor B

Alternate Names

VEGFB

Entrez Gene IDs

7423 (Human); 22339 (Mouse)

Gene Symbol

VEGFB

UniProt

P49766

Additional VEGF-B Products

Product Documents for Mouse VEGF-B 167/186 Antibody

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Product Specific Notices for Mouse VEGF-B 167/186 Antibody

For research use only

Citations for Mouse VEGF-B 167/186 Antibody

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