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Anti-TBP1/2 | TATA BINDING PROTEIN 1/2
Product no: AS23 4929
AS23 4929 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
329
€
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- Product Info
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Immunogen: KLH-conjugated peptide derived from protein sequence of Arabidopsis thaliana TBP1 and TBP2 UniProt: P28147 and P28148, TAIR:AT3G13445.1, AT1G55520.1 Host: Rabbit Clonality: Polyclonal Purity: Antigen affinity purified serum, in PBS pH 7.4 Format: Lyophilized Quantity: 50 µg Reconstitution: For reconstitution, add 50 µl, of sterile or deionized water. Storage: Store lyophilized/reconstituted at -20°C; once reconstituted, make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes. Tested applications: ChIP-qPCR chromatin immunoprecipitation (ChIP) combined with quantitative polymerase chain reaction (qPCR) Recommended dilution: 6 µg Expected | apparent MW: 22.4 kDa - Reactivity
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Confirmed reactivity: Arabidopsis thaliana Predicted reactivity: Arachis hypogaea, Brachypodium distachyon, Brassica napus, Cannabis sativa, Capsicum annuum, Glycine max, Hordeum vulgare, Malus domestica, Manihot esculenta, Medicago truncatula, Nicotania tabacum, Oryza sativa, Pisum sativum, Solanum lycopersicum, Solanum tuberosum, Sorghum bicolor, Spinacia oleracea, Theobroma cacao L, Triticum sp, Vitis vinifera, Zea mays
Species of your interest not listed? Contact usNot reactive in: No confirmed exceptions from predicted reactivity are currently known - Application Examples
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Chromatin extraction and immunoprecipitation was performed according to the aChIP protocol (https://doi.org/10.1038/s41477-024-01743-7). In particular, all the buffer formulations and wash schemes were preserved. In brief, ground in liquid nitrogen powder from 0.2g of green siliques of Arabidopsis thaliana, was crosslinked for 10 min in 4°C in 20ml of MC buffer(0.1M sucrose, 10mM sodium phosphate pH 7.0, 50mM NaCl) containing 1% formaldehyde. After stopping the crosslink with 0.125 M glycine, the pellet was resuspended in Z-1 buffer. Single washes using Z-2 and Z-3 were performed and the chromatin was resuspended in 2 ml of Z-4. Samples were sonicated using Bioruptor (Diagenode) apparatus for 26 cycles (30sec ON/30sec OFF; Mode: High). The average fragment size was around 500bp, estimated from agarose gel migration. After sonication and 10 min centrifugation, the soluble supernatant was collected and directly used for IP. Before, 12 µl of Protein-G-Dynabeads (Thermo Sci) were incubated in PBS supplemented with 0.5% BSA and with or without 6µg of anti-TBP antibodies (Agrisera) for 4 hrs in 4°C with constant rotation, prior to adding to chromatin. Immunoprecipitation reactions were set for overnight. Each chromatin sample was split to 2 IP reactions (TBP and NoAb, 600µl each) and 1 Input sample (60µl). On the next day, IP reactions were washed twice using Low Salt, once using High Salt followed by LiCl Wash solutions, then twice using TE buffer. DNA was recovered from IP and Input saples by 15 min incubation in 10% chelex suspension in 95°C, followed by proteinase K digestion (2h in 50°C with 2µl of Proteinase K) and inactivation (10 min in 95°C). qPCR was performed on IP samples, No Antibody controls and 1:10 diluted Input samples (1:100 - total dilution factor, relative to IP), using SYBR I qPCR mix (Roche).Courtesy of Dr. Lien Brzeźniak, Polish Academy of Science, Warsaw, Poland
- Additional Information
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Additional information (application): The peptide used to elicit this antibody is conserved in both isoforms: 1 and 2 of TATA Binding Protein. - Background
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Background: TBP1/2 (TATA BINDING PROTEIN 1/2) is a general transcription factor that functions at the core of the DNA-binding multiprotein factor TFIID which plays a role in the activation of eukaryotic genes transcribed by RNA polymerase II. Alternative names: TATA sequence-binding protein 1 (TBP-1), TATA-binding factor 1, TATA-box factor 1, Transcription initiation factor TFIID TBP-1 subunit.
- Protocols
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Agrisera Western Blot protocol and video tutorials
Protocols to work with plant and algal protein extracts
Agrisera Educational Poster Collection - Reviews:
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