Revision 1
#18950
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IP, ChIP
Reactivity:
H
Sensitivity:
Endogenous
MW (kDa):
280
Source/Isotype:
Rabbit IgG
UniProt ID:
#Q6N021
Entrez-Gene Id:
54790
Product Usage Information
For optimal ChIP results, use 20 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:100 |
Chromatin IP | 1:25 |
Storage
Specificity/Sensitivity
TET2 (D6B9Y) Rabbit mAb recognizes endogenous levels of total TET2 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human TET2 protein.
Background
Methylation of DNA at cytosine residues is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting, and mammalian development (1,2). 5-methylcytosine is a repressive epigenetic mark established de novo by two enzymes, DNMT3a and DNMT3b, and is maintained by DNMT1 (3, 4). 5-methylcytosine was originally thought to be passively depleted during DNA replication. However, subsequent studies have shown that Ten-Eleven Translocation (TET) proteins TET1, TET2, and TET3 can catalyze the oxidation of methylated cytosine to 5-hydroxymethylcytosine (5-hmC) (5). Additionally, TET proteins can further oxidize 5-hmC to form 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC), both of which are excised by thymine-DNA glycosylase (TDG), effectively linking cytosine oxidation to the base excision repair pathway and supporting active cytosine demethylation (6,7). TET2 is the most frequently mutated gene in myeloid dysplastic syndrome (MDS), a dysplasia of myeloid, megakaryocytic, and/or erythroid cell lineages, of which 30% progress to acute myeloid leukemia (AML) (8, 9). It is also mutated in diffuse large B-cell lymphoma (10). TET2 protein expression is often reduced in solid tumors such as prostate cancer, melanoma, and oral squamous cell carcinoma (11-13).
Background References
- Hermann, A. et al. (2004) Cell Mol Life Sci 61, 2571-87.
- Turek-Plewa, J. and Jagodziński, P.P. (2005) Cell Mol Biol Lett 10, 631-47.
- Okano, M. et al. (1999) Cell 99, 247-57.
- Li, E. et al. (1992) Cell 69, 915-26.
- Tahiliani, M. et al. (2009) Science 324, 930-5.
- He, Y.F. et al. (2011) Science 333, 1303-7.
- Ito, S. et al. (2011) Science 333, 1300-3.
- Langemeijer, S.M. et al. (2009) Nat Genet 41, 838-42.
- Yamazaki, J. et al. (2012) Epigenetics 7, 201-7.
- Asmar, F. et al. (2013) Haematologica 98, 1912-20.
- Nickerson, M.L. et al. (2013) Hum Mutat 34, 1231-41.
- Lian, C.G. et al. (2012) Cell 150, 1135-46.
- Jäwert, F. et al. (2013) Anticancer Res 33, 4325-8.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
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Revision 1
#18950
TET2 (D6B9Y) Rabbit mAb
Western blot analysis of extracts from DLD-1 or 293T cells, either wild type (+/+) or TET2 knockout (-/-), using TET2 (D6B9Y) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, DLD-1 and 293T (+/+) cells are positive for TET2 expression, while TET2 knockout (-/-) cells are negative.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293T cells and either TET2 (D6B9Y) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ZNF335 Promoter Primers #25946, human TAF12 exon1 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.