Revision 1

#25497Store at -20C

1 个试剂盒

(4 x 20 microliters)

Cell Signaling Technology

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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
JARID1A (D28B10) Rabbit mAb 3876 20 µl 200 kDa Rabbit IgG
JARID1B (E2X6N) Rabbit mAb 15327 20 µl 180 kDa Rabbit IgG
JARID1C (D29B9) Rabbit mAb 5361 20 µl 180 kDa Rabbit IgG
JARID1D (E4D4B) Rabbit mAb 78995 20 µl 190 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 JARID1/KDM5 Histone Demethylase Antibody Sampler Kit provides an economical means for detecting the JARID1/KDM5 family of proteins. The kit includes enough antibodies to perform two western blot experiments with each 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 antibodies.

Background

The methylation state of lysine residues in histone proteins is a major determinant for formation of active and inactive regions of the genome and is crucial for proper programming of the genome during development (1,2). Jumonji C (JmjC) domain-containing proteins represent the largest class of potential histone demethylase proteins (3). The JmjC domain can catalyze the demethylation of mono-, di-, and tri-methyl lysine residues via an oxidative reaction that requires iron and α-ketoglutarate (3). Based on homology, both humans and mice contain at least 30 such proteins, which can be divided into 7 separate families (3). The JARID (Jumonji/AT-rich interactive domain-containing protein) family contains four members: JARID1A (also RBP2 and RBBP2), JARID1B (also PLU-1), JARID1C (also SMCX), and JARID1D (also SMCY) (4). In addition to the JmJC domain, these proteins contain JmJN, BRIGHT, C5HC2 zinc-finger, and PHD domains, the latter of which binds to methylated histone H3 (Lys9) (4). All four JARID proteins demethylate di- and tri-methyl histone H3 Lys4; JARID1B also demethylates mono-methyl histone H3 Lys4 (5-7). JARID1A is a critical RB-interacting protein and is required for Polycomb-Repressive Complex 2 (PRC2)-mediated transcriptional repression during ES cell differentiation (8). A JARID1A-NUP98 gene fusion is associated with myeloid leukemia (9). JARID1B, which interacts with many proteins including c-Myc and HDAC4, may play a role in cell fate decisions by blocking terminal differentiation (10-12). JARID1B is overexpressed in many breast cancers and may act by repressing multiple tumor suppressor genes, including BRCA1 and HOXA5 (13,14). JARID1C has been found in a complex with HDAC1, HDAC2, G9a, and REST, which binds to and represses REST target genes in non-neuronal cells (7). JARID1C mutations are associated with X-linked mental retardation and epilepsy (15,16). JARID1D is uniquely localized to the Y chromosome, and functions as a tumor suppressor by repressing genes associated with cell invasiveness (17). JARID1D is frequently mutated in metastatic prostate tumors, and low JARID1D levels are associated with poor prognosis in prostate cancer patients (17).

  1. Kubicek, S. et al. (2006) Ernst Schering Res Found Workshop, 1-27.
  2. Lin, W. and Dent, S.Y. (2006) Curr Opin Genet Dev 16, 137-42.
  3. Klose, R.J. et al. (2006) Nat Rev Genet 7, 715-27.
  4. Benevolenskaya, E.V. (2007) Biochem Cell Biol 85, 435-43.
  5. Christensen, J. et al. (2007) Cell 128, 1063-76.
  6. Yamane, K. et al. (2007) Mol Cell 25, 801-12.
  7. Tahiliani, M. et al. (2007) Nature 447, 601-5.
  8. Pasini, D. et al. (2008) Genes Dev 22, 1345-55.
  9. van Zutven, L.J. et al. (2006) Genes Chromosomes Cancer 45, 437-46.
  10. Secombe, J. et al. (2007) Genes Dev 21, 537-51.
  11. Barrett, A. et al. (2007) Int J Cancer 121, 265-75.
  12. Dey, B.K. et al. (2008) Mol Cell Biol 28, 5312-27.
  13. Barrett, A. et al. (2002) Int J Cancer 101, 581-8.
  14. Lu, P.J. et al. (1999) J Biol Chem 274, 15633-45.
  15. Tzschach, A. et al. (2006) Hum Mutat 27, 389.
  16. Jensen, L.R. et al. (2005) Am J Hum Genet 76, 227-36.
  17. Li, N. et al. (2016) Cancer Res 76, 831-43.

Background References

    Trademarks and Patents

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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    仅供研究使用。不得用于诊断流程。
    Revision 1
    #25497

    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit

    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 1 Expand Image
    使用 Jarid1B (E2X6N) 兔单克隆抗体对 MCF7、NCCIT 和 COS-7 细胞系的提取物进行蛋白印迹分析。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 2 Expand Image
    使用 CUT&RUN Assay Kit #86652 对 K-562 细胞和 JARID1B (E2X6N) Rabbit mAb 进行 CUT&RUN 检测。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示了跨 GAPDH 基因的 JARID1B 结合。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 3 Expand Image
    使用 JARID1A (D28B10) Rabbit mAb 对来自不同细胞系的提取物进行蛋白质印迹分析。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 4 Expand Image
    使用 JARID1C (D29B9) Rabbit mAb 对 NCCIT 和 F9 细胞系提取物进行蛋白质印迹法分析。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 5 Expand Image
    一抗与靶标蛋白结合之后，与偶联 HRP 的二抗形成复合体。添加 LumiGLO®，在酶催化分解期间发光。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 6 Expand Image
    使用 JARID1D (E4D4B) Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 7 Expand Image
    对 MCF7 细胞提取物 Jarid1B 进行免疫沉淀。泳道 1 为 10% 输入，泳道 2 为兔 (DA1E) 单克隆抗体 IgG XP® 同型对照 #3900，泳道 3 为 Jarid1B (E2X6N) 兔单克隆抗体。使用 Jarid1B (E2X6N) 兔单克隆抗体进行蛋白印迹分析。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 8 Expand Image
    使用 CUT&RUN Assay Kit #86652 对 K-562 细胞和 JARID1B (E2X6N) Rabbit mAb 进行 CUT&RUN 检测。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在染色体 12（上图）内的 JARID1B 结合，包括 GAPDH 基因（下图）。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 9 Expand Image
    对 THP-1 细胞提取物的 JARID1D 蛋白进行免疫沉淀分析。泳道 1 为 10% input，泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900，泳道 3 为 JARID1D (E4D4B) Rabbit mAb。使用 JARID1D (E4D4B) Rabbit mAb 进行蛋白质印迹分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 10 Expand Image
    使用 SimpleChIP® 酶解染色质免疫共沉淀试剂盒（磁珠法） #9005 对 K562 细胞的交联染色质和 Jarid1B (E2X6N) 兔单克隆抗体 进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在 Jarid1B 的已知靶基因 GAPDH 内结合（参见包含 ChIP-qPCR 数据的其他结果图）。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 11 Expand Image
    使用 CUT&RUN Assay Kit #86652 对 K-562 细胞和 JARID1B (E2X6N) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362进行 CUT&RUN 检测。使用 SimpleChIP® Human γ-Actin Intron 3 Primers #5047 和 SimpleChIP® Human α Satellite Repeat Primers #4486 进行实时 PCR 来对富集的 DNA 进行定量。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。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 12 Expand Image
    使用 SimpleChIP® 酶解染色质免疫共沉淀试剂盒（磁珠法） #9005 对 K562 细胞的交联染色质和 Jarid1B (E2X6N) 兔单克隆抗体 进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在染色体12（上图）内的结合，包括 Jarid1B 的已知靶标基因 GAPDH（下图）（参见包含 ChIP-qPCR 数据的其他结果图）。
    JARID1/KDM5 Histone Demethylase Antibody Sampler Kit: Image 13 Expand Image
    使用 SimpleChIP® 酶解染色质免疫共沉淀试剂盒（磁珠法） #9005 对 K-562 细胞的交联染色质与 Jarid1B (E2X6N) 兔单克隆抗体或正常兔 IgG #2729 进行染色质免疫沉淀。使用 SimpleChIP® Human GAPDH Exon 1 Primers #5516、SimpleChIP® Human γ-Actin Intron 3 Primers #5047、SimpleChIP® Human DHFR Intron 1 Primers #7531 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。
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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    仅供研究使用。不得用于诊断流程。