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.

使用に関する制限

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Revision 9
#8238

p75NTR (D4B3) XP® Rabbit mAb

Western Blotting Image 1: p75NTR (D4B3) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from PC-3 cells, untreated (-) or ibuprofen-treated (2 mM, 24 hr) (+), and SW480 cells (-) using p75NTR (D4B3) XP® Rabbit mAb (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Immunoprecipitation Image 1: p75NTR (D4B3) XP® Rabbit mAb Expand Image
Immunoprecipitation of p75NTR from SW-480 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is p75NTR (D4B3) XP® Rabbit mAb. Western blot was performed using p75NTR (D4B3) XP® Rabbit mAb.
Immunofluorescence Image 1: p75NTR (D4B3) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse optic tract labeled with p75NTR (D4B3) XP® Rabbit mAb (green) and DAPI #8961 (blue).
Immunofluorescence Image 2: p75NTR (D4B3) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen developing mouse E12 spinal cord and somites labeled with p75NTR (D4B3) XP® Rabbit mAb (left, green), DyLight 554 Phalloidin #13054 (right, red), and DAPI #8961 (right, blue).
Immunofluorescence Image 1: p75NTR (D4B3) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of SK-N-MC cells using p75NTR (D4B3) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow Cytometry Image 1: p75NTR (D4B3) XP® Rabbit mAb Expand Image
Flow cytometric analysis of live DLD-1 cells (blue, negative) and SK-N-MC cells (green, positive) using p75NTR (D4B3) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.