Revision 4

#61702Store at -20C

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.
Applications:

WB, IP, ChIP, ChIP-seq, C&R

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

300

Source/Isotype:

Rabbit IgG

UniProt ID:

#O15047

Entrez-Gene Id:

9739

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 5 μL of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Chromatin IP 1:100
Chromatin IP-seq 1:100
CUT&RUN 1:100

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.

Specificity / Sensitivity

SET1A (D3V9S) Rabbit mAb recognizes endogenous levels of total SET1A protein. This antibody does not cross-react with other SET1/COMPASS proteins, including SET1B, MLL1, MLL2, MLL3, and MLL4.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro383 of human SET1A protein.

Background

The Set1 histone methyltransferase protein was first identified in yeast as part of the Set1/COMPASS histone methyltransferase complex, which methylates histone H3 at Lys4 and functions as a transcriptional co-activator (1). While yeast contain only one known Set1 protein, six Set1-related proteins exist in mammals: SET1A, SET1B, MLL1, MLL2, MLL3, and MLL4, all of which assemble into COMPASS-like complexes and methylate histone H3 at Lys4 (2,3). These Set1-related proteins are each found in distinct protein complexes, all of which share the common subunits WDR5, RBBP5, ASH2L, CXXC1 and DPY30. These subunits are required for proper complex assembly and modulation of histone methyltransferase activity (2-6). MLL1 and MLL2 complexes contain the additional protein subunit, menin (6). Like yeast Set1, all six Set1-related mammalian proteins methylate histone H3 at Lys4 (2-6). MLL translocations are found in a large number of hematological malignancies, suggesting that Set1/COMPASS histone methyltransferase complexes play a critical role in leukemogenesis (6).

  1. Miller, T. et al. (2001) Proc Natl Acad Sci USA 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.

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

WB: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

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 All: all species expected

Trademarks and Patents

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

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 4
#61702

SET1A (D3V9S) Rabbit mAb

Western Blotting Image 1: SET1A (D3V9S) Rabbit mAb Expand Image
使用 SET1A (D3V9S) Rabbit mAb 对 293T 和 HeLa 细胞的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: SET1A (D3V9S) Rabbit mAb Expand Image
对 293T 细胞提取物 SET1A 进行免疫沉淀。泳道 1 为10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 SET1A (D3V9S) Rabbit mAb。使用 SET1A (D3V9S) Rabbit mAb 进行蛋白质印迹分析。
Chromatin Immunoprecipitation Image 1: SET1A (D3V9S) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 HeLa 细胞的交联染色质和 SET1A (D3V9S) Rabbit mAb 进行染色质免疫沉淀。使用 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795 制成 DNA 库。本图显示在 GAPDH (SET1A 的一个已知靶标基因)内的结合作用(参见包含 ChIP-qPCR 数据的其他结果图)。欲知其他 ChIP-seq 情况,请下载产品说明书。
Chromatin Immunoprecipitation Image 2: SET1A (D3V9S) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 HeLa 细胞的交联染色质和 SET1A (D3V9S) Rabbit mAb 进行染色质免疫沉淀。使用 SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795 制成 DNA 库。结果图显示在染色体 12(上图)内的结合,包括 SET1A 的已知靶标基因 GAPDH(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: SET1A (D3V9S) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对 HeLa 细胞中提取的交联染色质,在加入 SET1A (D3V9S) Rabbit mAb 或Normal Rabbit IgG #2729 后,进行染色质免疫沉淀。使用 SimpleChIP® Human β-Actin Promoter Primers #13653、SimpleChIP® Human GAPDH Promoter Primers #4471 和 SimpleChIP® Human α Satellite Repeat Primers #4486,通过实时 PCR 对富集的 DNA 进行定量分析。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。
CUT and RUN Image 1: SET1A (D3V9S) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 ,以 HeLa 细胞和 SET1A (D3V9S) Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示结合作用遍及 PRKAA1 基因, SET1A 的已知靶标基因(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 2: SET1A (D3V9S) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 ,以 HeLa 细胞和 SET1A (D3V9S) Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示在染色体 5(上图)内的结合,包括 SET1A 的已知靶标基因 PRKAA1(下图)(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 3: SET1A (D3V9S) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,以 HeLa 细胞和 SET1A (D3V9S) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN。使用人 PRKAA1 启动子引物、人 KANSL1L 启动子引物和人 PAD14 启动子引物,通过实时 PCR 对富集的 DNA 定量。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。