Revision 6
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:

ChIP, ChIP-seq, C&R, C&T

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9H9S0

Entrez-Gene Id:

79923

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 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.

The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

Application Dilution
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 1:50
CUT&Tag 1:50

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

Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) detects endogenous levels of total nanog protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of human nanog protein.

Background

Nanog is a homeodomain-containing transcription factor that is essential for the maintenance of pluripotency and self renewal in embryonic stem cells (1). Nanog expression is controlled by a network of factors including Sox2 and the key pluripotency regulator Oct-4 (1). Recent advances in somatic cell reprogramming have utilized viral expression of combinations of transcription factors including nanog, Oct-4, Sox2, KLF4, c-Myc, and LIN28 (2,3).

  1. Kim, J. et al. (2008) Cell 132, 1049-61.
  2. Takahashi, K. et al. (2007) Nat Protoc 2, 3081-9.
  3. Yu, J. et al. (2007) Science 318, 1917-20.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN C&T: CUT&Tag

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.
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

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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 6
#5232

Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated)

Chromatin Immunoprecipitation Image 1: Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 对 NCCIT 细胞的交联染色质与 Oct-4A (C30A3C1) Rabbit mAb (ChIP Formulated) #5677 或 Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) 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。经发现 Oct-4 和 Nanog 在染色质上互相关联。结果图显示在 POU5F1/Oct-4 内结合 Oct-4 和 Nanog，POU5F1/Oct-4 已知是 Oct-4 和 Nanog 的一个靶标基因 （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: Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003，对 NCCIT 细胞的交联染色质与 10 μl Oct-4A (C30A3C1) Rabbit mAb (ChIP Formulated) #5677 或 Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) 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。经发现 Oct-4 和 Nanog 在染色质上互相关联。图中显示通过染色体结合 Oct-4 和 Nanog6（上），其中包括 POU5F1/Oct-4（下），一种 Oct-4 和 Nanog 的已知靶标基因 （参见含 ChIP-qPCR 数据的附图）。
Chromatin Immunoprecipitation Image 3: Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 对 NTERA-2 细胞的交联染色质和 Nanog、Oct-4 和 Sox2 抗体、或 Normal Rabbit IgG 进行染色质免疫沉淀。使用 human Nanog 启动子引物、SimpleChIP® Human Oct-4 Promoter Primers #4641、SimpleChIP® Human Sox2 Promoter Primers #4649 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。
Chromatin Immunoprecipitation Image 4: Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 对 NCCIT 细胞的交联染色质和 Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated)、或 Normal Rabbit IgG #2729 进行染色质免疫沉淀。使用 SimpleChIP® Human Oct-4 Promoter Primers #4641、SimpleChIP® Human Sox2 Promoter Primers #4649 和 SimpleChIP® Human α Satellite Repeat Primers #4486，通过实时 PCR 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。
CUT and RUN Image 1: Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) Expand Image
使用 CUT&RUN Assay Kit #86652 对 NCCIT 细胞和 Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) 进行 CUT&RUN 分析。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示在 Nanog 的已知靶基因 Oct-4/POU5F1 内的结合（参见包含 CUT&RUN-qPCR 数据的附加图）。
CUT and RUN Image 2: Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) Expand Image
使用 CUT&RUN Assay Kit #86652 对 NCCIT 细胞和 Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) 进行 CUT&RUN 分析。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示在染色体 6（上图）内的结合，包括 Nanog 的已知靶标基因 Oct-4/POU5F1（下图）（参见包含 CUT&RUN-qPCR 数据的其他结果图）。
CUT and RUN Image 3: Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) Expand Image
使用CUT&RUN Assay Kit #86652 对€‚ NCCIT 细胞和 Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN 分析。使用 SimpleChIP® Human Oct-4 Promoter Primers #4641, SimpleChIP® Human Sox2 Promoter Primers #4649 和人 ITM2A 启动子引物，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 Tag Image 1: Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) Expand Image
CUT&Tag was performed with NCCIT cells and Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated), 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 SOX2, a known target gene of Nanog (see our ChIP-qPCR figure).
CUT and Tag Image 2: Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated) Expand Image
CUT&Tag was performed with NCCIT cells and Nanog (D73G4) XP® Rabbit mAb (ChIP Formulated), 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 3 (upper), including SOX2 (lower), a known target gene of Nanog (see our ChIP-qPCR figure).
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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。