Revision 9

#8238Store 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, IF-F, IF-IC, FC-L

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

75

Source/Isotype:

Rabbit IgG

UniProt ID:

#P08138

Entrez-Gene Id:

4804

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Frozen) 1:1600 - 1:3200
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:3200
Flow Cytometry (Live) 1:200 - 1:800

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 #46333.

Specificity / Sensitivity

p75NTR (D4B3) XP® Rabbit mAb recognizes endogenous levels of total p75NTR protein. Nonspecific cytoplasmic staining is observed in fixed frozen mouse spleen and colon by immunofluorescence.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg198 of human p75NTR protein. This antibody is predicted to bind the extracellular amino-terminal region of p75NTR protein.

Background

The p75 neurotrophin receptor (p75NTR), a member of the TNF receptor superfamily, is distinguished by multiple cysteine-rich ligand-binding domains, a single transmembrane sequence, and a noncatalytic cytoplasmic domain (1). p75NTR displays paradoxical functions when acting alone or with other receptor proteins. Working in concert with Trk receptors, p75NTR recognizes neurotrophins and transmits trophic signals into the cell. Both p75NTR and TrkA are required to activate PI3K-Akt signaling, while TrkA can individually activate the MAP kinase pathway. In contrast, p75NTR, possibly through JNK, ensures appropriate apoptosis of injured neurons and improperly targeted neonatal neurons (2).

The p75NTR protein undergoes sequential cleavage similar to APP and Notch. First, α-secretase removes the p75NTR ectodomain, eliminating ligand-mediated signaling. At this point, the membrane-tethered cleavage product can still fine-tune Trk-mediated trophic actions. γ-secretase cleaves within the transmembrane domain to liberate the cytoplasmic tail from its membrane anchor and allow the p75NTR intracellular domain to translocate to the nucleus (3,4).

  1. Chao, M.V. (2003) Nat. Rev. Neurosci. 4, 299-309.
  2. Nykjaer, A. et al. (2005) Curr. Opin. Neurobiol. 15, 49-57.
  3. Kanning, K.C. et al. (2003) J. Neurosci. 23, 5425-5436.
  4. Jung, K.M. et al. (2003) J. Biol. Chem. 278, 42161-42169.

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 IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-L: Flow Cytometry (Live)

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

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 9
#8238

p75NTR (D4B3) XP® Rabbit mAb

Western Blotting Image 1: p75NTR (D4B3) XP® Rabbit mAb Expand Image
使用 p75NTR (D4B3) XP® Rabbit mAb (上图)和 Akt (pan) (C67E7) Rabbit mAb #4691(下图)对未经处理 (-) 或已经 ibuprofen(2 mM,24 小时)(+) 处理的 PC-3 细胞和 SW480 细胞 (-) 的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: p75NTR (D4B3) XP® Rabbit mAb Expand Image
对 SW-480 细胞提取物中的 p75NTR 进行免疫沉淀分析。泳道 1 为 10% 输入,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 p75NTR (D4B3) XP® Rabbit mAb。使用 p75NTR (D4B3) XP® Rabbit mAb 进行蛋白质印迹分析。
Immunofluorescence Image 1: p75NTR (D4B3) XP® Rabbit mAb Expand Image
对使用 p75NTR (D4B3) XP® Rabbit mAb(绿色)和 DAPI #8961(蓝色)标记的固定冷冻的小鼠视神经束进行共聚焦免疫荧光分析。
Immunofluorescence Image 2: p75NTR (D4B3) XP® Rabbit mAb Expand Image
对使用 p75NTR (D4B3) XP® Rabbit mAb(左图,绿色)、DyLight 554 Phalloidin #13054(右图,红色)和 DAPI #8961(右图,蓝色)标记的固定冷冻的发育小鼠 E12 脊髓和体节进行共聚焦免疫荧光分析。
Immunofluorescence Image 1: p75NTR (D4B3) XP® Rabbit mAb Expand Image
使用 p75NTR (D4B3) XP® Rabbit mAb (绿色)对 SK-N-MC 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。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: p75NTR (D4B3) XP® Rabbit mAb Expand Image
使用 p75NTR (D4B3) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对活性 DLD-1 细胞(蓝色,阴性)和 SK-N-MC 细胞(绿色,阳性)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。