Revision 8

#2258Store 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, W-S, IP, IHC-P, IF-F, IF-IC, FC-FP, C&R

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

42

Source/Isotype:

Rabbit IgG

UniProt ID:

#P17947

Entrez-Gene Id:

6688

Product Information

Product Usage Information

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:100
Immunofluorescence (Frozen) 1:200 - 1:400
Immunofluorescence (Immunocytochemistry) 1:200 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:200
CUT&RUN 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.

For a carrier free (BSA and azide free) version of this product see product #45150.

Specificity / Sensitivity

This antibody detects endogenous levels of total PU.1 protein. The antibody does not cross react with other Ets family members.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Pig

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence of human PU.1 protein.

Background

PU.1 is a member of the Ets family of transcription factors and activates target genes through the purine-rich PU-box (1). PU.1 plays a pivotal role in the differentiation of myeloid cells and lymphocytes and is expressed in several hematopoietic cells including B lymphocytes, macrophages, neutrophils, mast cells, early erythroid cells, and megakaryocytes (1,2). The concentration of PU.1 is critical for both the determination of hematopoietic cell lineage and the regulation of differentiation versus stem cell proliferation (3,4). In addition, PU.1 activity is influenced by phosphorylation and interactions with other hematopoietic transcription factors. Phosphorylation of PU.1 at Ser146 by CK2 promotes binding to IRF4 and synergistic activation through the immunoglobulin κ 3' enhancer (5). Treatment of pro-B cells with IL-3 leads to phosphorylation of PU.1 at Ser140, resulting in increased PU.1 activity and activation of the anti-apoptotic gene MCL-1 (6). GATA1 binding blocks PU.1 activity during erythroid cell development (7). Overexpression of PU.1 resulting from proviral insertion during Friend virus infection can induce erythroleukemia, while reduced expression has been associated with acute myeloid leukemia (8).

  1. Lloberas, J. et al. (1999) Immunol Today 20, 184-9.
  2. Klemsz, M.J. et al. (1990) Cell 61, 113-24.
  3. Dahl, R. and Simon, M.C. (2003) Blood Cells Mol Dis 31, 229-33.
  4. DeKoter, R.P. and Singh, H. (2000) Science 288, 1439-41.
  5. Pongubala, J.M. et al. (1993) Science 259, 1622-5.
  6. Wang, J.M. et al. (2003) Mol Cell Biol 23, 1896-909.
  7. Zhang, P. et al. (1999) Proc Natl Acad Sci U S A 96, 8705-10.
  8. Moreau-Gachelin, F. et al. (1988) Nature 331, 277-80.

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 W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) 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.
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 8
#2258

PU.1 (9G7) Rabbit mAb

Western Blotting Image 1: PU.1 (9G7) Rabbit mAb Expand Image
使用 PU.1 (9G7) Rabbit mAb 对 RAW、THP1 和 p388D1 细胞的提取物进行蛋白质印迹分析。
Western Blotting Image 1: PU.1 (9G7) Rabbit mAb Expand Image
Simple Western™ analysis of lysates (0.1 mg/mL) from THP-1 cells using PU.1 (9G7) Rabbit mAb #2258. 虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
No image available
Immunohistochemistry Image 1: PU.1 (9G7) Rabbit mAb Expand Image
使用 PU.1 (9G7) Rabbit mAb #2258 对石蜡包埋的 4T1 同基因小鼠肿瘤细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: PU.1 (9G7) Rabbit mAb Expand Image
在有对照肽(左图)或 PU.1 Blocking Peptide #1036(右图)的情况下,使用 PU.1 (9G7) Rabbit mAb 对石蜡包埋的非霍奇金淋巴瘤细胞进行免疫组织化学分析。
Immunofluorescence Image 1: PU.1 (9G7) Rabbit mAb Expand Image
使用 PU.1 (9G7) 兔单克隆抗体(黄色)和 GFAP (GA5) 小鼠单克隆抗体(红色)对正常小鼠脑细胞进行共聚焦免疫荧光分析。在用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭游离二抗结合位点后,该组织随后用 ASC (D2W8U) Rabbit mAb (Mouse Specific)(Alexa 488 Conjugate)(绿色)标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Immunofluorescence Image 1: PU.1 (9G7) Rabbit mAb Expand Image
使用 PU.1 (9G7) Rabbit mAb(绿色)对 Raw 细胞进行共聚焦免疫荧光分析。使用 DY-554 phalloidin(红色)标记肌动蛋白纤丝。
Flow Cytometry Image 1: PU.1 (9G7) Rabbit mAb Expand Image
使用 PU.1 (9G7) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线),对 MCF7 细胞(蓝色)和 THP1 细胞(绿色)进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。
CUT and RUN Image 1: PU.1 (9G7) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 U-937 细胞和 PU.1 (9G7) 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。该图显示在 TOMM40L 基因内结合。
CUT and RUN Image 2: PU.1 (9G7) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 U-937 细胞和 PU.1 (9G7) 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。该图显示在染色体 1(上图)内的结合,包括 TOMM40L 基因(下图)。
CUT and RUN Image 3: PU.1 (9G7) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 U-937 细胞和 PU.1 (9G7) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN 检测。使用 human ANKRD44 intron 21 primers、human CTSS exon 1 primers 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。