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877-616-CELL (2355)

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Support:

877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
G9a/EHMT2 (C6H3) Rabbit Monoclonal Antibody330620 µl160,180 kDaRabbit IgG
ESET (C1C12) Rabbit Monoclonal Antibody219620 µl180 kDaRabbit IgG
ASH2L (D93F6) Rabbit Monoclonal Antibody501920 µl80, 65 kDaRabbit IgG
RBBP5 (D3I6P) Rabbit Monoclonal Antibody1317120 µl70 kDaRabbit IgG
SET7/SET9 Antibody281320 µl48 kDaRabbit
SMYD2 (D14H7) Rabbit Monoclonal Antibody973420 µl49 kDaRabbit IgG
SUV39H1 (D11B6) Rabbit Monoclonal Antibody872920 µl48 kDaRabbit IgG
SET8 (C18B7) Rabbit Monoclonal Antibody299620 µl43 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

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 two western blot experiments per primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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).

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using RBBP5 (D3I6P) Rabbit mAb.
undefined Image 1: Lysine Methyltransferase Antibody Sampler Kit
CUT&Tag was performed with HeLa cells and RBBP5 (D3I6P) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across CDKN2C, a known target gene of RBBP5 (see additional figure containing ChIP-qPCR data).
undefined Image 2: Lysine Methyltransferase Antibody Sampler Kit
CUT&RUN was performed with HeLa cells and RBBP5 (D3I6P) Rabbit mAb #13171, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across CDKN2C gene, a known target gene of RBBP5 (see additional figure containing CUT&RUN-qPCR data).
undefined Image 3: Lysine Methyltransferase Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of cell lysates from MCF7 and 293 cells using ESET (C1C12) Rabbit mAb.
undefined Image 4: Lysine Methyltransferase Antibody Sampler Kit
Western blot analysis of cell lysates from HeLa, NIH/3T3, C6 and COS cells using SET7/SET9 Antibody.
undefined Image 5: Lysine Methyltransferase Antibody Sampler Kit
Western blot analysis of extracts from HCT116 and RAW cells using SET8 (C18B7) Rabbit mAb.
undefined Image 6: Lysine Methyltransferase Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from HeLa and 293 cells using G9a/EHMT2 (C6H3) Rabbit mAb.
undefined Image 7: Lysine Methyltransferase Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using ASH2L (D93F6) XP® Rabbit mAb.
undefined Image 8: Lysine Methyltransferase Antibody Sampler Kit
CUT&RUN was performed with F9 cells and ASH2L (D93F6) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across the Tfeb gene.
undefined Image 9: Lysine Methyltransferase Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
undefined Image 10: Lysine Methyltransferase Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using SUV39H1 (D11B6) Rabbit mAb.
undefined Image 11: Lysine Methyltransferase Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using SMYD2 (D14H7) Rabbit mAb.
undefined Image 12: Lysine Methyltransferase Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunoprecipitation of RBBP5 from C2C12 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or RBBP5 (D3I6P) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using RBBP5 (D3I6P) Rabbit mAb.
undefined Image 13: Lysine Methyltransferase Antibody Sampler Kit
CUT&RUN was performed with HeLa cells and RBBP5 (D3I6P) Rabbit mAb #13171, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome #1 (upper), including CDKN2C (lower), a known target gene of RBBP5 (see additional figure containing CUT&RUN-qPCR data).
undefined Image 14: Lysine Methyltransferase Antibody Sampler Kit
CUT&Tag was performed with HeLa cells and RBBP5 (D3I6P) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 1 (upper), including CDKN2C (lower), a known target gene of RBBP5 (see additional figure containing ChIP-qPCR data).
undefined Image 15: Lysine Methyltransferase Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of MCF-7 cells using ESET (C1C12) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
undefined Image 16: Lysine Methyltransferase Antibody Sampler Kit
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with 0.5% Triton X-100 for 10 minutes prior to fixation according to Tardat et al. (2007) J. Cell Biol. 179, 1413-26 (right), using SET8 (C18B7) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
undefined Image 17: Lysine Methyltransferase Antibody Sampler Kit
Confocal immunofluorescent analysis of HeLa cells using G9a/EHMT2 (C6H3) Rabbit mAb (green). Actin filaments have been labeled with DY-555 phalloidin (red).
undefined Image 18: Lysine Methyltransferase Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Confocal imunofluorescent analysis of HeLa cells using ASH2L (D93F6) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
undefined Image 19: Lysine Methyltransferase Antibody Sampler Kit
CUT&RUN was performed with F9 cells and ASH2L (D93F6) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 17 (upper), including the Tfeb gene (lower).
undefined Image 20: Lysine Methyltransferase Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either RBBP5 (D3I6P) or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CDKN1B Promoter Primers #11951, human CDKN2C promoter 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.
undefined Image 21: Lysine Methyltransferase Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
CUT&RUN was performed with HeLa cells and RBBP5 (D3I6P) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human CDKN2C promoter 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.
undefined Image 22: Lysine Methyltransferase Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from K562 cells and either G9a/EHMT2 (C6H3) or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ZNF19 Intron 3 Primers #98428, human VRK3 intron 11 primers, and SimpleChIP® Human GAPDH Exon 1 Primers #5516. 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.
undefined Image 23: Lysine Methyltransferase Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.