Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766)
Key features and details
- Rabbit polyclonal to TATA binding protein TBP - Nuclear Loading Control and ChIP Grade
- Suitable for: IP, ChIP, IHC-P, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade
See all TATA binding protein TBP primary antibodies -
Description
Rabbit polyclonal to TATA binding protein TBP - Nuclear Loading Control and ChIP Grade -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ChIP HumanIHC-P HumanIP HumanWB MouseHuman
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Immunogen
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Images
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ChIP - Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766)Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab63766 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach for active loci and Taqman approach for inactive loci). Primers and probes are located in the first kb of the transcribed region.
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Western blot - Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766)All lanes : Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766) at 1 µg/ml
Lane 1 : Testis (Mouse) Tissue Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : Testis (Rat) Tissue Lysate
Lane 4 :Hep G2 nuclear extract lysate (ab14660)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 38 kDa
Observed band size: 38,45 kDa why is the actual band size different from the predicted?
Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766) This image is courtesy of an anonymous Abreviewab63766 staining TAT binding protein TBP in human infantile fibromatosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 3 hours at room temperature; antigen retrieval was by heat mediation in Tris pH9. Samples were incubated with primary antibody (1/100 in TBS + 1% BSA + 1% FBS) for 16 hours. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.
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Immunoprecipitation - Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766)TATA binding protein TBP was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to TATA binding protein TBP and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab63766.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 45kDa: TATA binding protein TBP. -
Western blot - Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766)All lanes : Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766) at 1 µg/ml
Lane 1 :Recombinant Human TATA binding protein TBP (ab81897) at 0.1 µg
Lane 2 :Recombinant Human TATA binding protein TBP (ab81897) at 0.01 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 38 kDa
Exposure time: 10 seconds -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766)IHC image of TATA binding protein TBP staining in Human breast ductal carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab63766, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Western blot - Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766)All lanes : Anti-TATA binding protein TBP antibody - Nuclear Loading Control and ChIP Grade (ab63766) at 1 µg/ml
Lane 1 : HeLa Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 38 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Additional bands at: 60 kDa. We are unsure as to the identity of these extra bands.
