Revision 1

#17944Store 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

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

38

SOURCE:

Rabbit

UniProt ID:

#P06748

Entrez-Gene Id:

4869

Product Information

Product Usage Information

For optimal ChIP results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 × 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Chromatin IP 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

NPM1 (C Mutant Specific) Antibody recognizes endogenous levels of total NPM1 C mutant protein. This antibody may also detect a band around 220 kDa that likely corresponds to pentameric NPM1 C mutant protein. This antibody does not cross-react with wild-type NPM1 protein.

Species Reactivity:

Human

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the mutated carboxy terminus of human NPM1 C mutant protein. Antibodies are purified by peptide affinity chromatography.

Background

Nucleophosmin (NPM1; also known as NPM, B23, numatrin, or NO38) is an abundant phosphoprotein primarily found in nucleoli. It has been implicated in several distinct cellular functions, including assembly and transport of ribosomes, cytoplasmic/nuclear trafficking, regulation of DNA polymerase α activity, centrosome duplication, and molecular chaperoning activities (1,2). The NPM1 gene is also known for its fusion with the anaplastic lymphoma kinase (ALK) receptor tyrosine kinase. The NPM1 portion contributes to transformation by providing a dimerization domain, which results in activation of the fused kinase (3,4). NPM1 is also the most frequently mutated gene in acute myeloid leukemia (AML) and accounts for nearly 30% of all cases (5). These AML subtypes, classified as NPM1-mutated AML, are characterized by mutations in NPM1’s C-terminus that disrupt its nucleolar localization sequence and cause mislocalization from the nucleolus to the cytoplasm (6). This cytoplasmic form of NPM1, commonly referred to as NPM1c, is exclusive to myeloid malignancies and is not found in other forms of cancer (7). These mutations are always heterozygous, and NPM1c functions in a dominant negative fashion by dimerizing with wild-type NPM1 and recruiting it to the cytoplasm (6,8). Interestingly, NPM1 mutations alone are not sufficient to drive leukemogenesis, and further research is required to fully elucidate the impact of these mutations on disease progression (9).

  1. Okuda, M. et al. (2000) Cell 103, 127-40.
  2. Takemura, M. et al. (1999) J Biochem 125, 904-9.
  3. Morris, S.W. et al. (1994) Science 263, 1281-4.
  4. Bischof, D. et al. (1997) Mol Cell Biol 17, 2312-25.
  5. Falini, B. et al. (2005) N Engl J Med 352, 254-66.
  6. Falini, B. et al. (2006) Blood 107, 4514-23.
  7. Zarka, J. et al. (2020) Genes (Basel) 11, .
  8. den Besten, W. et al. (2005) Cell Cycle 4, 1593-8.
  9. Vassiliou, G.S. et al. (2011) Nat Genet 43, 470-5.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP

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.
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 1
#17944

NPM1 (C Mutant Specific) Antibody

Western Blotting Image 1: NPM1 (C Mutant Specific) Antibody Expand Image
使用 NPM1 (C Mutant Specific) Antibody(上图)、NPM1 Antibody #3542(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对 OCI-AML3 和 HeLa 细胞的提取物进行蛋白质印迹法分析。HeLa 细胞中 NPM1 C 突变蛋白的阴性表达与已发表的观察结果一致。
Immunoprecipitation Image 1: NPM1 (C Mutant Specific) Antibody Expand Image
对 OCI-AML3 细胞提取物的 NPM1 C 蛋白突变体进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Normal Rabbit IgG #2729,泳道 3 为 NPM1 (C Mutant Specific) Antibody。使用 NPM1 (C Mutant Specific) Antibody 进行蛋白质印迹法分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Chromatin Immunoprecipitation Image 1: NPM1 (C Mutant Specific) Antibody Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 OCI-AML3 细胞中的交联染色质 和 NPM1 (C Mutant Specific) Antibody 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀分析。使用人 CDKN2C 外显子 2 引物、人 IRX5 外显子 3 引物、人 ISL2 内含子 3 引物和 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。