Explore
Validate
Learn
Western blot
Gel shiftAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- MA1-25427 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TBP Monoclonal Antibody (1TB18)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- MA1-25427 detects TATA binding protein TBP from most mammal species samples.
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 1TB18
- Vial size
- 50 µL
- Concentration
- 1 mg/mL
- Storage
- 4° C
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of TBP was achieved by transfecting HeLa with TBP specific siRNAs (Silencer® select Product # s13826, s13827). Western blot analysis (Fig. a) was performed using nuclear enriched extracts from the TBP knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2), and untransfected cells (lane 1). The blot was probed with TBP Monoclonal Antibody (1TB18) (Product # MA1-25427, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in the histogram (Fig. b). The decrease in signal upon siRNA mediated knockdown confirms that the antibody is specific to TBP.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-TBP Monoclonal Antibody (1TB18) (Product # MA1-25427) and a 41kDa band corresponding to TBP was observed across all cell lines tested, except NIH/3T3 and mouse and rat testes tissues where a 37kDa band was observed. Nuclear enriched extracts (30 µg lysate) of K-562 (Lane 1), HeLa (Lane 2), A-431 (Lane 3), NIH/3T3 (Lane 4), HEL 92.1.7 (Lane 5), Mouse Testis (Lane 6), Rat Testis (Lane 7) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of TBP was performed using 70% confluent log phase HeLa cells. 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 TBP Monoclonal Antibody (1TB18) (Product # MA1-25427) 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 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 dilution). Panel d represents the merged image showing predominant nuclear localization. Panel e represents control cells with no primary antibody to assess the background. The images were captured at 60X magnification.
