Product Pathways - Chromatin Regulation / Epigenetics

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Lysine Methyltransferase Antibody Sampler Kit #8694

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
G9a/EHMT2 (C6H3) Rabbit mAb #3306	40 µl	W IF-IC	H M R Mk (B) (Pg) (Hr)	140, 165	Rabbit IgG
ESET (C1C12) Rabbit mAb #2196	40 µl	W IP IF-IC	H Mk	180	Rabbit IgG
ASH2L (D93F6) XP® Rabbit mAb #5019	40 µl	W IP IF-IC	H M R Mk (Dm)	80, 65	Rabbit IgG
RBBP5 Antibody #8037	40 µl	W	H M R Mk (C) (X) (B) (Pg) (Hr)	67	Rabbit
SET7/SET9 Antibody #2813	40 µl	W IF-IC	H M R Mk	48	Rabbit
SMYD2 (D14H7) Rabbit mAb #9734	40 µl	W IP	H M R Mk (Hm) (B) (Dg) (Hr)	49	Rabbit IgG
SUV39H1 (D11B6) Rabbit mAb #8729	40 µl	W IP	H M R Mk (B)	48	Rabbit IgG
SET8 (C18B7) Rabbit mAb #2996	40 µl	W IF-IC	H M R Mk (B) (Pg) (Hr)	43, 50	Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  Dm=D. melanogaster  X=Xenopus  B=Bovine  Dg=Dog  Pg=Pig  Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in this kit recognizes only the specific target protein and does not cross-react with other family members.

Description

The Lysine Methyltransferase Antibody Sampler Kit provides a fast and economical means to evaluate endogenous levels of lysine methyltransferases. The kit contains enough primary antibody to perform four western blot experiments per primary antibody.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human ASH2L protein, the carboxy terminus of human ESET protein, the carboxy terminus of human G9a/EHMT2 protein, human SET8 protein, residues surrounding Val414 of human SMYD2 protein, or residues surrounding Asp380 of human SUV39H1 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val407 of human RBBP5 protein or the human SET7/SET9 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

SET domain-containing proteins are potential histone methyltransferases (HMTases), which are classified into subgroups by their putative substrate specificities. Histone H3 Lys9 (H3-K9) methyltransferase group genes include Suv39h1, Suv39h2, G9a, G9a related protein (GLP) and SETDB1/ESET (1). The H3-K9 methylation mark plays an important role as a binding site for the chromo-containing protein, resulting in chromatin compaction and heterochromatin generation (2). Histone H3-K4 methylation is exclusively associated with actively transcribed genes (2). The first H3-K4 methylase complex, COMPASS, was identified in the yeast S. cerevisiae and consists of Set1/KMT2 and seven other polypeptides, Cps60-Cps15 (2). Set1/KMT2 functions within COMPASS and is capable of mono-, di-, and trimethylating H3-K4 (2). There are several Set1 related proteins in mammals including WDR5, RBBP5, ASH2L, CXXC1, and DPY30 (2,3). SET7/SET9 is a member of the SET domain-containing family that can specifically methylate H3-K4, Lys189 of the TAF10, a member of the TFIID transcription factor complex, and Lys372 of the p53 tumor suppressor protein (4-6). SET domain-containing lysine methyltransferase 8 (SET8), also known as PR/SET domain-containing protein 7 (PR/SET7), is a single-subunit enzyme that mono-methylates histone H4-K20, preferably on nucleosomal substrates (7-9). SET and MYND domain-containing protein 2 (SMYD2), also known as lysine methyltransferase protein 3C (KMT3C), functions to repress transcription by interacting with the Sin3A repressor complex and methylating H3-K36 (10). SMYD2 also methylates H3-K4 through interaction with HSP90α, and methylates p53 at Lys370 to repress p53-mediated transcriptional activation and apoptosis (11,12).

1. Tachibana, M. et al. (2005) Genes Dev 19, 815-26.
2. Shilatifard, A. (2008) Curr Opin Cell Biol 20, 341-8.
3. Lee, J.H. et al. (2007) J Biol Chem 282, 13419-28.
4. Nishioka, K. et al. (2002) Genes Dev 16, 479-89.
5. Kouskouti, A. et al. (2004) Mol Cell 14, 175-82.
6. Chuikov, S. et al. (2004) Nature 432, 353-60.
7. Fang, J. et al. (2002) Curr Biol 12, 1086-99.
8. Xiao, B. et al. (2005) Genes Dev 19, 1444-54.
9. Couture, J.F. et al. (2005) Genes Dev 19, 1455-65.
10. Brown, M.A. et al. (2006) Mol Cancer 5, 26.
11. Abu-Farha, M. et al. (2008) Mol Cell Proteomics 7, 560-72.
12. Huang, J. et al. (2006) Nature 444, 629-32.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

2196:

Immunofluorescence, Immunoprecipitation, Western Blotting

2813:

Immunofluorescence, Western Blotting

2996:

Immunofluorescence, Western Blotting

3306:

Immunofluorescence, Western Blotting

5019:

Immunofluorescence, Immunoprecipitation, Western Blotting

8037:

Western Blotting

8729:

Immunoprecipitation, Western Blotting

9734:

Immunoprecipitation, Western Blotting

Companion Products

• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 7071 Phototope^®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7075 Anti-biotin, HRP-linked Antibody
• 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
• 9998 BSA
• 9999 Nonfat Dry Milk

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For Research Use Only. Not For Use In Diagnostic Procedures.