Revision 1

#25501Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
SET1A (D3V9S) Rabbit mAb 61702 20 µl 300 kDa Rabbit IgG
SET1B (D1U5D) Rabbit mAb 44922 20 µl 320 kDa Rabbit IgG
MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen) 14689 20 µl 300 kDa Rabbit IgG
MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) 14197 20 µl 180 kDa Rabbit IgG
MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen) 63735 20 µl 80 kDa Rabbit IgG
WDR5 (D9E1I) Rabbit mAb 13105 20 µl 37 kDa Rabbit IgG
WDR82 (D2I3B) Rabbit mAb 99715 20 µl 30 kDa Rabbit IgG
Menin (D45B1) XP® Rabbit mAb 6891 20 µl 68 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

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

Description

The SET1/COMPASS Antibody Sampler Kit provides an economical means of detecting SET1/COMPASS proteins using control antibodies against SET1A, SET1B, MLL1, MLL2, WDR5, WDR82, and Menin. This kit contains enough primary antibodies to perform at least two western blot experiments.

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

The Set1 histone methyltransferase protein was first identified in yeast as part of the Set1/COMPASS histone methyltransferase complex, which methylates histone H3 on lysine 4 and functions as a transcriptional co-activator (1). While yeast contain only one known Set1 protein, mammals contain six Set1-related proteins: SET1A, SET1B, MLL1, MLL2, MLL3 and MLL4, all of which methylate histone H3 on lysine 4 (2,3). These Set1-related proteins are each found in distinct protein complexes, all of which share the common core structural subunits WDR5, RBBP5 and ASH2L (2-6). WDR82 is a core subunit specific to SET1A and SET1B complexes, while Menin is a core subunit specific to the MLL complexes (4,5,7).

Like yeast Set1, all six Set1-related mammalian proteins methylate histone H3 on lysine 4 (2-6). SET1A, SET1B, MLL1 and MLL2 mediate di- and tri-methylation of histone H3 Lys4 at gene promoters to facilitate transcription activation. MLL3 and MLL4 function primarily to mono-methylate histone H3 Lys4 at gene enhancers. MLL1 and MLL2 function as master regulators of both embryogenesis and hematopoiesis, and are required for proper expression of Hox genes (8-10). MLL1 is a large approximately 4000 amino acid protein that is cleaved by the Taspase 1 threonine endopeptidase to form N-terminal (MLL1-N) and C-terminal MLL1 (MLL1-C) fragments, both of which are subunits of the functional MLL1/COMPASS complex (11,12). MLL1 translocations are found in a large number of hematological malignancies, suggesting that Set1 histone methyltransferase complexes play a critical role in leukemogenesis (6). Like MLL1, MLL2 is also a large, approximately 2700 amino acid protein that is cleaved by the Taspase 1 threonine endopeptidase to form N-terminal (MLL2-N) and C-terminal (MLL2-C) fragments, both of which are subunits of the functional MLL2/COMPASS complex. MLL2 has also been implicated as a modulator of hematological malignancies (13). MLL3 and MLL4 proteins are not cleaved by Taspase 1.

  1. Miller, T. et al. (2001) Proc Natl Acad Sci U S A 98, 12902-7.
  2. Shilatifard, A. (2008) Curr Opin Cell Biol 20, 341-8.
  3. Tenney, K. and Shilatifard, A. (2005) J Cell Biochem 95, 429-36.
  4. Lee, J.H. and Skalnik, D.G. (2005) J Biol Chem 280, 41725-31.
  5. Lee, J.H. et al. (2007) J Biol Chem 282, 13419-28.
  6. Hughes, C.M. et al. (2004) Mol Cell 13, 587-97.
  7. Yokoyama, A. et al. (2004) Mol Cell Biol 24, 5639-49.
  8. Eissenberg, J.C. and Shilatifard, A. (2010) Dev Biol 339, 240-9.
  9. Smith, E. et al. (2011) Genes Dev 25, 661-72.
  10. Denissov, S. et al. (2014) Development 141, 526-37.
  11. Takeda, S. et al. (2006) Genes Dev 20, 2397-409.
  12. Yokoyama, A. et al. (2002) Blood 100, 3710-8.
  13. Chen, Y. et al. (2017) Cancer Cell 31, 755-770.e6.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a 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.

    使用に関する制限

    法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    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
    #25501

    SET1/COMPASS Antibody Sampler Kit

    SET1/COMPASS Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extracts from various cell lines using WDR5 (D9E1I) Rabbit mAb.
    SET1/COMPASS Antibody Sampler Kit: Image 2 Expand Image
    CUT&RUN was performed with NCCIT cells and WDR5 (D9E1I) Rabbit mAb, using CUT&RUN Assay Kit #86652. The DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across HOXA genes.
    SET1/COMPASS Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from MLL1 wild-type (WT) and knockout (KO) mouse embryonic fibroblasts (MEF) using MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). MLL1 WT and KO MEF were kindly provided by Dr. Ali Shilatifard of Northwestern University.
    SET1/COMPASS Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from various cell lines using MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen).
    SET1/COMPASS Antibody Sampler Kit: Image 5 Expand Image
    CUT&RUN was performed with NCCIT cells and MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen), 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 ELOC, a known target gene of MLL1 (see additional figure containing CUT&RUN-qPCR data).
    SET1/COMPASS Antibody Sampler Kit: Image 6 Expand Image
    CUT&Tag was performed with NCCIT cells and MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen), 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 ELOC gene.
    SET1/COMPASS Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from 293T, F9, and COS-7 cell lines using SET1B (D1U5D) Rabbit mAb.
    SET1/COMPASS Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from 293T and HeLa cells using SET1A (D3V9S) Rabbit mAb.
    SET1/COMPASS Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various cell lines using MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen) (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, HCT 116 cells are negative for MLL2 expression.
    SET1/COMPASS Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from various cell lines using Menin (D45B1) XP® Rabbit mAb.
    SET1/COMPASS Antibody Sampler Kit: Image 11 Expand Image
    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.
    SET1/COMPASS Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from HCT 116, NIH/3T3, and COS-7 cell lines using WDR82 (D2I3B) Rabbit mAb.
    SET1/COMPASS Antibody Sampler Kit: Image 13 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and WDR5 (D9E1I) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NANOG, a known target gene of WDR5 (see additional figure containing ChIP-qPCR data).
    SET1/COMPASS Antibody Sampler Kit: Image 14 Expand Image
    CUT&RUN was performed with NCCIT cells and WDR5 (D9E1I) Rabbit mAb, using CUT&RUN Assay Kit #86652. The DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across HOXA genes (upper) and HOXD genes (lower).
    SET1/COMPASS Antibody Sampler Kit: Image 15 Expand Image
    Western blot analysis of extracts from various cell lines using MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen).
    SET1/COMPASS Antibody Sampler Kit: Image 16 Expand Image
    CUT&RUN was performed with NCCIT cells and MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen), using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 8 (upper), including ELOC (lower), a known target gene of MLL1 (see additional figure containing CUT&RUN-qPCR data).
    SET1/COMPASS Antibody Sampler Kit: Image 17 Expand Image
    CUT&Tag was performed with NCCIT cells and MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen), 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 8 (upper), including ELOC gene (lower).
    SET1/COMPASS Antibody Sampler Kit: Image 18 Expand Image
    Immunoprecipitation of SET1B from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is SET1B (D1U5D) Rabbit mAb. Western blot analysis was performed using SET1B (D1U5D) Rabbit mAb.
    SET1/COMPASS Antibody Sampler Kit: Image 19 Expand Image
    Immunoprecipitation of SET1A from 293T cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is SET1A (D3V9S) Rabbit mAb. Western blot analysis was performed using SET1A (D3V9S) Rabbit mAb.
    SET1/COMPASS Antibody Sampler Kit: Image 20 Expand Image
    Immunoprecipitation of MLL2/KMT2B from NCCIT cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen). Western blot analysis was performed using MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen).
    SET1/COMPASS Antibody Sampler Kit: Image 21 Expand Image
    Confocal immunofluorescent analysis of COS-7 cells using Menin (D45B1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    SET1/COMPASS Antibody Sampler Kit: Image 22 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and WDR5 (D9E1I) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 12 (upper), including NANOG (lower), a known target gene of WDR5 (see additional figure containing ChIP-qPCR data).
    SET1/COMPASS Antibody Sampler Kit: Image 23 Expand Image
    CUT&RUN was performed with NCCIT cells and either WDR5 (D9E1I) 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 TAF6L promoter primers and human ZNF644 promoter primers. 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.
    SET1/COMPASS Antibody Sampler Kit: Image 24 Expand Image
    Confocal immunofluorescent analysis of MEF wild-type (left, positive) and MEF MLL (-/-) (right, negative) cells using MLL (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). MLL1 WT and KO MEF were kindly provided by Dr. Ali Shilatifard of Northwestern University.
    SET1/COMPASS Antibody Sampler Kit: Image 25 Expand Image
    CUT&RUN was performed with NCCIT cells and either MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen) 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 RTN4 promoter, human ELOC promoter, 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.
    SET1/COMPASS Antibody Sampler Kit: Image 26 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and SET1A (D3V9S) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across GAPDH, a known target gene of SET1A (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
    SET1/COMPASS Antibody Sampler Kit: Image 27 Expand Image
    Confocal immunofluorescent analysis of A549 cells (left, positive) and HCT 116 cells (right, negative) using MLL2/KMT2B (D6X2E) Rabbit mAb (Carboxy-terminal Antigen) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
    SET1/COMPASS Antibody Sampler Kit: Image 28 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either WDR5 (D9E1I) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human nanog promoter primers, SimpleChIP® Human Oct-4 Promoter Primers #4641, 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.
    SET1/COMPASS Antibody Sampler Kit: Image 29 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and SET1A (D3V9S) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 12 (upper), including GAPDH (lower), a known target gene of SET1A (see additional figure containing ChIP-qPCR data).
    SET1/COMPASS Antibody Sampler Kit: Image 30 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either SET1A (D3V9S) Rabbit mAb 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 β-Actin Promoter Primers #13653, SimpleChIP® Human GAPDH Promoter Primers #4471, 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.
    SET1/COMPASS Antibody Sampler Kit: Image 31 Expand Image
    CUT&RUN was performed with HeLa cells and SET1A (D3V9S) Rabbit mAb, 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 PRKAA1 gene, a known target gene of SET1A (see additional figure containing CUT&RUN-qPCR data).
    SET1/COMPASS Antibody Sampler Kit: Image 32 Expand Image
    CUT&RUN was performed with HeLa cells and SET1A (D3V9S) Rabbit mAb, 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 5 (upper), including PRKAA1 (lower), a known target gene of SET1A (see additional figure containing CUT&RUN-qPCR data).
    SET1/COMPASS Antibody Sampler Kit: Image 33 Expand Image
    CUT&RUN was performed with HeLa cells and either SET1A (D3V9S) 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 PRKAA1 promoter primers, human KANSL1L promoter primers, and human PAD14 promoter primers. 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.
    Image Gallery
    Learn more about how we get our images
    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.