Revision 1
#25501
Store at -20C
SET1/COMPASS Antibody Sampler Kit
1 Kit
(8 x 20 microliters)
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
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
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.
Background References
- Miller, T. et al. (2001) Proc Natl Acad Sci U S A 98, 12902-7.
- Shilatifard, A. (2008) Curr Opin Cell Biol 20, 341-8.
- Tenney, K. and Shilatifard, A. (2005) J Cell Biochem 95, 429-36.
- Lee, J.H. and Skalnik, D.G. (2005) J Biol Chem 280, 41725-31.
- Lee, J.H. et al. (2007) J Biol Chem 282, 13419-28.
- Hughes, C.M. et al. (2004) Mol Cell 13, 587-97.
- Yokoyama, A. et al. (2004) Mol Cell Biol 24, 5639-49.
- Eissenberg, J.C. and Shilatifard, A. (2010) Dev Biol 339, 240-9.
- Smith, E. et al. (2011) Genes Dev 25, 661-72.
- Denissov, S. et al. (2014) Development 141, 526-37.
- Takeda, S. et al. (2006) Genes Dev 20, 2397-409.
- Yokoyama, A. et al. (2002) Blood 100, 3710-8.
- Chen, Y. et al. (2017) Cancer Cell 31, 755-770.e6.
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.
Limited Uses
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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.
Revision 1
Western blot analysis of extracts from various cell lines using WDR5 (D9E1I) Rabbit mAb.
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.
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.
Revision 1
Western blot analysis of extracts from various cell lines using MLL1 (D2M7U) Rabbit mAb (Amino-terminal Antigen).
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).
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.
Revision 1
Western blot analysis of extracts from 293T, F9, and COS-7 cell lines using SET1B (D1U5D) Rabbit mAb.
Western blot analysis of extracts from 293T and HeLa cells using SET1A (D3V9S) Rabbit mAb.
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.
Revision 1
Western blot analysis of extracts from various cell lines using Menin (D45B1) XP® Rabbit mAb.
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.
Western blot analysis of extracts from HCT 116, NIH/3T3, and COS-7 cell lines using WDR82 (D2I3B) Rabbit mAb.
Revision 1
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).
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).
Western blot analysis of extracts from various cell lines using MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen).
Revision 1
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).
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).
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.
Revision 1
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.
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).
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).
Revision 1
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).
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.
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.
Revision 1
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.
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.
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).
Revision 1
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.
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).
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.
Revision 1
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).
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).
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.