MA5-12184

antibody from Invitrogen Antibodies

Targeting: VEGFA VEGF, VEGF-A, VPF

Antibody data

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Validation data

Product number: MA5-12184 - Provider product page
Provider: Invitrogen Antibodies
Product name: VEGF Monoclonal Antibody (VG1)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-12184 targets Vascular Endothelial Growth Factor in IF, IHC (P) and WB applications and shows reactivity with Human samples.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: VG1
Vial size: 500 µL
Concentration: 0.2 mg/mL
Storage: 4° C

VEGFA protein structure - MA5-12184 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Vascular Endothelial Growth Factor using Vascular Endothelial Growth Factor Monoclonal Antibody (Product # MA5-12184) on VEGF Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Vascular Endothelial Growth Factor (VEGF) was performed by loading 1 µg of indicated recombinant VEGF isoforms, and 10 µL of PageRuler PlusPrestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product # 88518) using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. VEGF was detected at 14 kD, 19 kD and 22-30 kD using a VEGF monoclonal antibody (Product # MA5-12184) at a concentration of 1 µg/mL in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG (Fc) secondary antibody (Product # 31437) at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-VEGF Monoclonal Antibody (VG1) (Product # MA5-12184) and a 52 kDa band corresponding to VEGF was observed across cell line tested in Figure a and 20 and 40 kDa bands in recombinant VEGF protein in Figure b. Whole cell extracts (30 µg lysate) of HACAT cells and HACAT cells treated with 0.5 mM H2O2 for 30 minutes followed by 1X-PTI for 4 hours in Figure a and recombinant VEGF protein (250 ng and 25 ng) in Figure b were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of VEGF was performed using DU 145 cells and DU 145 cells treated with 1X-PTI for 4 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with VEGF Monoclonal Antibody (VG1) (Product # MA5-12184) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of DU 145 PTI-treated cells showing induction of VEGF protein in cytoplasm (golgi apparatus). Panel e represents the merged image of untreated DU 145 cells, that shows lower expression for VEGF protein. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of VEGF was performed using HACAT cells and HACAT cells treated with 0.5 mM H2O2 for 30 minutes followed by 1X-PTI for 4 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with VEGF Monoclonal Antibody (VG1) (Product # MA5-12184) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing induction of VEGF expression in cytoplasm in PTI-treated HACAT cells. Panel e represents untreated HACAT cells showing lower VEGF expression. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Vascular Endothelial Growth Factor (VEGF, green) in Hela cells. The cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 in PBS, and blocked with 3% Blocker BSA (Product # 37525) for 30 minutes at room temperature. Cells were stained with (left panel) or without (right panel) a VEGF polyclonal antibody (Product # MA5-12184) at a dilution of 1:40 for at least 1 hour at room temperature, and then incubated with a Dylight 488 goat anti-Mouse IgG secondary antibody at a dilution of 1:1000 for 45 minutes at room temperature. F-actin (both panels, red) was stained by Dylight 554 Phalloidin (Product # 21834) and nuclei (both panels, blue) were stained with DAPI (Product # 46190). Images were taken at 60X magnification.