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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [2]
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- Product number
- MA5-15276 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- FGF2 Monoclonal Antibody (2H5G2C11)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-15276 targets FGF2 in IHC and WB applications and shows reactivity with Human samples.
- Antibody clone number
- 2H5G2C11
- Concentration
- Conc. Not Determined
Submitted references c-Myc-mediated repression of miR-15-16 in hypoxia is induced by increased HIF-2α and promotes tumor angiogenesis and metastasis by upregulating FGF2.
Xue G, Yan HL, Zhang Y, Hao LQ, Zhu XT, Mei Q, Sun SH
Oncogene 2015 Mar 12;34(11):1393-406
Oncogene 2015 Mar 12;34(11):1393-406
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of FGF2 using a FGF2 monoclonal antibody (Product # MA5-15276) against a truncated FGF2 recombinant protein.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of FGF2 was achieved by transfecting BJ with FGF2 specific siRNAs (Silencer® select Product # s223530, s5128). Western blot analysis (Fig. a) was performed using whole cell extracts from the FGF2 knockdown cells (Lane 3), non-targeting scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). Considering that FGF2 is a secretory protein and detection with total lysates is low in western blot application, untransfected, scrambled, FGF2 siRNA transfected cells were treated with PTI post-transfection to allow for their accumulation within the cells. The blot was probed with FGF2 Monoclonal Antibody (2H5G2C11) (Product # MA5-15276, 1:1500 dilution ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:8000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to FGF2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-FGF2 Monoclonal Antibody (2H5G2C11) (Product # MA5-15276) and a 22kDa band corresponding to FGF2 was observed in the fibroblast cell line BJ but not in MCF7 and T-47D upon treatment with the protein secretory blocker. Whole cell extracts (30 µg lysate) of BJ treated with PTI (1X for 4 hours) (Lane 1), untreated BJ (Lane 2), T-47D treated with PTI (1X for 4 hours) (Lane 3), untreated T-47D (Lane 4), MCF7 treated with PTI (1X for 4 hours) (Lane 5) and untreated MCF7 (Lane 6) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:8000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). FGF2, a protein of the fibroblast growth factor family, is secreted in high abundance by fibroblast cell lines like BJ. Entrapment of FGF2 using PTI increases intracellular accumulation in BJ but not in T-47D and MCF7 [10.1158/1078-0432.CCR-10-2727].
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of FGF2 was performed using 80% confluent log phase BJ and T-47D, control and PTI treated, cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with FGF2 Monoclonal Antibody (2H5G2C11) (Product # MA5-15276) at 1:100 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), (1:2500 dilution), for 45 minutes at room temperature (Panel a,e,i: Green). Nuclei (Panel b,f,j: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c,g,k: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of untreated BJ cells showing faint staining for FGF2 that is enhanced upon PTI treatment (Panel h). Panel l represents T-47D cells showing no staining for FGF2 upon PTI treatment. The images were captured at 60X magnification. FGF2 follows type I unconventional secretory pathway where it undergoes self-oligomerization and interacts with the inner and outer leaflets of the plasma membrane for its release. The punctate pattern of FGF2 seen in panel d and h is similar to that shown by Steringer et.al. of the intermediates and oligomers of FGF2 for membrane insertion.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded human rectum adenocarcinoma tissue showing cytoplasmic localization using FGF2 monoclonal antibody (Product # MA5-15276) followed with DAB staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded human rectum adenocarcinoma tissue showing cytoplasmic localization using FGF2 monoclonal antibody (Product # MA5-15276) followed with DAB staining.
