Revision 6

#2880Store 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, IHC-P, IF-IC, FC-FP, ChIP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

78 to 82

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q12778

Entrez-Gene Id:

2308

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Immunocytochemistry) 1:50 - 1:200
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:200
Chromatin IP 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 #29336.

Specificity / Sensitivity

FoxO1 (C29H4) Rabbit mAb detects endogenous levels of total FoxO1 protein. The antibody does not detect exogenously expressed family members FoxO3a or FoxO4.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a GST-fusion protein corresponding to carboxy-terminal residues of human FoxO1.

Background

The Forkhead family of transcription factors is involved in tumorigenesis of rhabdomyosarcoma and acute leukemias (1-3). Within the family, three members (FoxO1, FoxO4, and FoxO3a) have sequence similarity to the nematode orthologue DAF-16, which mediates signaling via a pathway involving IGFR1, PI3K, and Akt (4-6). Active forkhead members act as tumor suppressors by promoting cell cycle arrest and apoptosis. Increased expression of any FoxO member results in the activation of the cell cycle inhibitor p27 Kip1. Forkhead transcription factors also play a part in TGF-β-mediated upregulation of p21 Cip1, a process negatively regulated through PI3K (7). Increased proliferation results when forkhead transcription factors are inactivated through phosphorylation by Akt at Thr24, Ser256, and Ser319, which results in nuclear export and inhibition of transcription factor activity (8). Forkhead transcription factors can also be inhibited by the deacetylase sirtuin (SirT1) (9).

  1. Anderson, M.J. et al. (1998) Genomics 47, 187-99.
  2. Galili, N. et al. (1993) Nat Genet 5, 230-5.
  3. Borkhardt, A. et al. (1997) Oncogene 14, 195-202.
  4. Nakae, J. et al. (1999) J Biol Chem 274, 15982-5.
  5. Rena, G. et al. (1999) J Biol Chem 274, 17179-83.
  6. Guo, S. et al. (1999) J Biol Chem 274, 17184-92.
  7. Seoane, J. et al. (2004) Cell 117, 211-23.
  8. Arden, K.C. (2004) Mol Cell 14, 416-8.
  9. Yang, Y. et al. (2005) EMBO J 24, 1021-32.

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 IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) 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.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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 6
#2880

FoxO1 (C29H4) Rabbit mAb

Western Blotting Image 1: FoxO1 (C29H4) Rabbit mAb Expand Image
使用 FoxO1 (C29H4) Rabbit mAb(上图)和 FoxO3a (75D8) Rabbit mAb #2497(下图)对 293T 细胞(野生型 (+/+) 或 FoxO1 (-/-))的提取物进行蛋白质印迹分析。
Western Blotting Image 2: FoxO1 (C29H4) Rabbit mAb Expand Image
使用 FoxO1 (C29H4) Rabbit mAb,对 IGROV-1 和 COS-7 细胞提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: FoxO1 (C29H4) Rabbit mAb Expand Image
对 C6 提取物 FoxO1 进行免疫沉淀。泳道 1 为 10% 输入,泳道 2 为 FoxO1 (C29H4) Rabbit mAb,泳道 3 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900。使用 FoxO1 (C29H4) Rabbit mAb 进行蛋白印迹分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Immunohistochemistry Image 1: FoxO1 (C29H4) Rabbit mAb Expand Image
使用 FoxO1 (C29H4) Rabbit mAb 对石蜡包埋的人类结肠进行免疫组织化学分析。
Immunohistochemistry Image 2: FoxO1 (C29H4) Rabbit mAb Expand Image
使用 FoxO1 (C29H4) Rabbit mAb 对石蜡包埋的人类肺癌进行免疫组织化学分析。
Immunohistochemistry Image 3: FoxO1 (C29H4) Rabbit mAb Expand Image
使用 FoxO1 (C29H4) Rabbit mAb,对石蜡包埋的人类淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 4: FoxO1 (C29H4) Rabbit mAb Expand Image
使用 FoxO1 (C29H4) Rabbit mAb,对用 LY294002(左)或 胰岛素(右)处理的石蜡包埋的 IGROV-1 细胞沉淀物,进行免疫组织化学分析。请注意,Akt 激活后 FoxO1 的胞质定位。
Immunohistochemistry Image 5: FoxO1 (C29H4) Rabbit mAb Expand Image
使用 FoxO1 (C29H4) Rabbit mAb,对石蜡包埋的 SKOV-3 异种移植组织进行免疫组织化学分析。
Immunofluorescence Image 1: FoxO1 (C29H4) Rabbit mAb Expand Image
使用 FoxO1 (C29H4) Rabbit mAb(绿色),对用 LY294002(左)或胰岛素(右)处理的 IGROV-1 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。蓝色伪彩 = DRAQ5 (DNA 荧光染料)。
Flow Cytometry Image 1: FoxO1 (C29H4) Rabbit mAb Expand Image
使用 FoxO1 (C29H4) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对 HL-60 细胞(蓝色)和 IGROV-1 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 作为二抗。
Chromatin Immunoprecipitation Image 1: FoxO1 (C29H4) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对小鼠肝组织的交联染色质与 FoxO1 (C29H4) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀分析。使用 SimpleChIP® Mouse Pdk4 Promoter Primers #31195、mouse Elk4 upstream primers 和 SimpleChIP® Mouse RPL30 Intron 2 Primers #7015 进行实时 PCR 来对富集的 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。