Revision 3
#36449
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IP, IF-IC, FC-FP
Reactivity:
M
Sensitivity:
Endogenous
MW (kDa):
280
Source/Isotype:
Rabbit IgG
UniProt ID:
#Q4JK59
Entrez-Gene Id:
214133
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Immunofluorescence (Immunocytochemistry) | 1:400 - 1:1600 |
Flow Cytometry (Fixed/Permeabilized) | 1:100 |
Storage
Specificity/Sensitivity
TET2 (D6C7K) Rabbit mAb recognizes endogenous levels of total TET2 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val1640 of Mouse 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 IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)
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
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Limited Uses
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Revision 3
#36449
TET2 (D6C7K) Rabbit mAb
Western blot analysis of extracts from mES, F9, and NIH/3T3 cell lines using TET2 (D6C7K) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, mES and F9 cells are positive for TET2 expression, while NIH/3T3 cells are negative for TET2 expression.
Immunoprecipitation of TET2 from mES cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TET2 (D6C7K) Rabbit mAb. Western blot analysis was performed using TET2 (D6C7K) Rabbit mAb.
Confocal immunofluorescent analysis of F9 cells (left, positive) and NIH/3T3 cells (right, negative) using TET2 (D6C7K) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Revision 3
#36449
TET2 (D6C7K) Rabbit mAb
Flow cytometric analysis of C2C12 cells (blue) and mES cells (green) using TET2 mouse (D6C7K) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alex Fluor® 488 Conjugate) #4412 was used as a secondary antibody.