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ChIPAb+ TATA Binding Protein (TBP) - ChIP Validated Antibody and Primer Set
Description:
ChIPAb+ TATA Binding Protein (TBP) - ChIP Validated Antibody and Primer Set
Trade Name:
Upstate (Millipore)
Product Overview:
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.
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Specificity:
Recognizes TATA Binding Protein (TBP).
Molecular Weight:
~42 kDa
Immunogen:
Full length recombinant human TBP.
Isotype:
IgG1
Background Information:
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).
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Species Reactivity:
Chicken
Human
Mouse
Xenopus
Fish
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Species Reactivity Note:
Human, mouse, chicken, xenopus and zebrafish. Does not react with Drososphila.
Reactivity with other species has not been confirmed.
Non-Reactive Species:
Drosophila
Application Notes:
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.
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Control:
Includes negative control mouse ascites and primers specific for human GAPDH promoter.
Quality Assurance:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from He |