Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Immunocytochemistry [3]
- Flow cytometry [1]
- Other assay [1]
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Validation data
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- Product number
- MA3-078 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TAF15 Monoclonal Antibody (8TA-2B10)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- MA3-078 detects TATA-Binding Protein (TBP) Associated Factor 15 from human, mouse, rat, canine, and non-human primate samples. The MA3-078 immunogen is the RNA- binding region (RNP) of human TAFII68 as a GST-fusion protein (from aa 175 to 414).
- Reactivity
- Human, Mouse, Rat, Canine
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 8TA-2B10
- Vial size
- 50 µL
- Concentration
- Conc. Not Determined
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of TAF15 in 3T3 cells. The cells were fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a TAF15 mouse monoclonal antibody (Product # MA3-078) at a dilution of 1:250 in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:1000 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of TAF15 in NRK cells. The cells were fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a TAF15 mouse monoclonal antibody (Product # MA3-078) at a dilution of 1:250 in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:1000 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of TAF15 in HepG2 cells. The cells were fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a TAF15 mouse monoclonal antibody (Product # MA3-078) at a dilution of 1:250 in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:1000 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of TAF15 was done on HeLa cells. Cells were fixed, permeabilized and stained with a TAF15 mouse monoclonal antibody (Product # MA3-078, red histogram) at a dilution of 1:100. After incubation of the primary antibody on ice for an hour, the cells were stained with a Goat anti-Mouse IgG (H+L) Secondary Antibody, DyLight 650 conjugate (Product # 84545) at a dilution of 1:50 for at least 30 minutes on ice. A representative 10,000 cells were acquired for each sample. The black histogram represents unstained control cells and the blue histogram represents no-primary-antibody control.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- RNA immunoprecipitation (RIP) western of TAF15 was performed on K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 µg of TAF15 monoclonal antibody (Product # MA3-078) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti-mouse coated Dynabeads (Product # 11202D), washed extensively, and eluted with NuPAGE™ LDS Sample Buffer (Product # NP0007). Samples were resolved onto NuPAGE™ 4-12% Bis-Tris gel (Product # NP0335BOX). Lanes 1 and 3 are input and lanes 2 and 4 are IP. Proteins were transferred to PVDF membrane (Product # IB23001). Membrane was blocked in 5% milk. Target was detected using a TAF15 monoclonal antibody (Product # MA3-078) at a dilution of 1:2000, followed by a 1:4000 dilution of secondary antibody. Chemiluminescent detection was performed using ECL Western Blotting Substrate (Product # 32106). Data courtesy of the Yeo lab as part of the ENCODE project (www.encodeproject.org).