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Merck

17-10098

Sigma-Aldrich

ChIPAb+ TATA Binding Protein (TBP) - ChIP Validated Antibody and Primer Set

ascites fluid, from mouse

Synonym(s):

TATA BINDING PROTEIN, TATA-box-binding protein, TATA-box factor, TATA-binding factor, TATA sequence-binding protein, Transcription initiation factor TFIID TBP subunit

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.52

Key Documents

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biological source

mouse

Quality Level

100

antibody form

ascites fluid

clone

monoclonal

species reactivity

human, zebrafish, mouse, Xenopus, chicken, fish

should not react with

Drosophila

manufacturer/tradename

ChIPAb+
Upstate®

technique(s)

ChIP: suitable
immunoprecipitation (IP): suitable
western blot: suitable

isotype

IgG1

NCBI accession no.

NP_003185

UniProt accession no.

P20226

shipped in

dry ice

Gene Information

human ... TBP(6908)

Related Categories

General description

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ TATA Binding Protein (TBP) set includes the TATA Binding Protein (TBP) antibody, a negative control mouse ascites, and qPCR primers which amplify a 166 bp region of human GAPDH promoter. The TATA Binding Protein (TBP) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of TATA Binding Protein (TBP)-associated chromatin.
TATA-box Binding Protein (TBP) is a general transcription factor that functions at the core of the DNA-binding multiprotein factor TFIID. Binding of TFIID to the TATA box is the initial transcriptional step of the pre-initiation complex (PIC). TBP plays a role in the activation of eukaryotic genes transcribed by RNA polymerase II and is a component of the transcription factor SL1/TIF-IB complex, which is involved in the assembly of the PIC (preinitiation complex) during RNA polymerase I-dependent transcription (Unitprot).
~42 kDa

Immunogen

Full length recombinant human TBP.

Application

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either Negative Ascites or 2 µL Anti-TATA Binding Protein (TBP) and the Magna ChIP® G Kit (Cat. # 17-611).
Successful immunoprecipitation of TBP associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH promoter as a positive locus, and GAPDH coding primers as a negative locus (Please see figures).
Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP G (Cat. # 17-409) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:
Representative lot data.
HeLa lysates were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-TATA Binding Protein (TBP) (1:1000 dilution).
Proteins were visualized using a secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).

Immunohistochemistry:
A 1:1,000-1:5,000 dilution of a previous lot was used in immunohistochemistry.

Immunocytochemistry:
A 1:1,000-1:5,000 dilution of a previous lot was used in immunocytochemistry.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Transcription Factors
The ChIPAb+ TATA Binding Protein (TBP) set includes the TATA Binding Protein (TBP) antibody, a negative control mouse ascites & qPCR primers which amplify a 166 bp region of human GAPDH promoter.

Biochem/physiol Actions

Reactivity with other species has not been confirmed.
Recognizes TATA Binding Protein (TBP).

Packaging

25 assays per set. Recommended use: ~2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).

Physical form

Anti-TATA Binding Protein (TBP) (mouse monoclonal). One vial containing 50 μL ascites containing 0.05% sodium azide. Store at -20°C.

Negative Ascites (mouse). One vial containing 50 µL of mouse ascites with 0.05% sodium azide. Store at -20°C.

ChIP Primers, GAPDH promoter. One vial containing 75 μL of 5 μM each primer specific for human GAPDH. Store at -20°C.
FOR: TAC TAG CGG TTT TAC GGG CG
REV: TCG AAC AGG AGG AGC AGA GAG CGA
Unpurified

Preparation Note

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Analysis Note

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either Negative Ascites or 2 µL Anti-TATA Binding Protein (TBP) and the Magna ChIP® G Kit (Cat. # 17-611).
Successful immunoprecipitation of TATA Binding Protein (TBP)-associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH promoter (Please see figures).

Please refer to the EZ-Magna ChIP G (Cat. # 17-409) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Control
Includes negative control mouse ascites and primers specific for human GAPDH promoter.

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage Class Code

10 - Combustible liquids

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Advance your Epigenetics Research

Epigenetics describes heritable changes in gene expression caused by non-genetic mechanisms. Epigenetic regulation allows a cell to vary its response based on its biological and environmental contexts. Epigenetic changes can effect transcriptional and post-transcriptional regulation via mechanisms such as histone modification, chromatin and nucleosome remodeling, DNA methylation, and small and non-coding RNA-mediated regulation. These mechanisms, in cooperation with transcription factors and other nucleic acid-binding proteins, regulate gene expression. Epigenetic mechanisms of gene regulation impacts diverse areas of research—from agriculture to human health. Common epigenetic assays such as chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) rely on high quality antibodies that recognize specific epigenetic modifications for accurate results. EMD Millipore offers over 100 ChIPAb+™ and RIPAb+™ validated antibody kits that are quality tested on ChIP/RIP assays and are conveniently provided with control qPCR primers and negative control antibodies to ensure first time ChIP/RIP success.

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