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NGFR

IHC of NGFR on an FFPE Brain Tissue

Description

NGFR (Nerve Growth Factor Receptor), also termed p75 or CD271, is the low-affinity NGFR (LNGFR) which binds NGF and other neurotrophins, including BDNF, NT3 and NT4/5 with similar low-affinity. NGFR p75 is a 75 kD transmembrane glycoprotein that is mainly expressed in Schwann cells and neurons and in a variety of non-neuronal cells. NGFR p75 is necessary for regulating neuronal growth, migration, differentiation and cell death during development of the central and peripheral nervous system. NGFR p75 plays a central role in the regulation of cell number by apoptosis in the developing CNS. During early development, activation of NGFR p75 by NGF induces apoptotic cell death in some neuronal cells, probably through activation of the sphingomyelinase/ceramide pathway, the ICE-like proteases and the JNK pathway. CD271 has recently been described as being expressed in mesenchymal stem cells (bone marrow stromal cells).


NGFR is expressed not only in sympathetic and sensory neurons, but also in various neural crest cell or tumor derivatives such as melanocytes, Melanomas, Neuroblastomas, Pheochromocytomas, Neuro“ bromas, and neurotized nevi (Type C melanocytes). It is now apparent that expression of NGFR is ubiquitous and not limited to the nervous system, being expressed in mature non-neural cells such as perivascular cells, dental pulp cells, lymphoid follicular dendritic cells, basal epithelium of oral mucosa and hair follicles, prostate basal cells and myoepithelial cells. Studies in Prostate and Urothelial Cancer suggest that NGFR may act as a tumor suppressor, negatively regulating cell growth and proliferation. NGFR labels the myoepithelial cells of breast ducts and intralobular “ broblasts of breast ducts and, thus, aids in the diagnosis of malignancy in the breast.

Antibody Type
Mouse Monoclonal
Clone
NGFR/c10
Isotype
IgG1
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Breast, CNS Tumors
Storage
Store at 2°-8°C
Stability
2 years

For long-term storage of the concentrated antibody, it is recommended that aliquots of the antibody be frozen at -20°C in glycerol 50% (frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Dilute using an antibody diluent such as ImmunoDetector Protein Block/Antibody Diluent (BSB 0040 and BSB 0041) or ImmunoDNA Background Blocker (BSB 0103-BSB 0107).

Presentation

NGFR is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.

Availability
Catalog No.
Antibody Type
Dilution
Volume/QTY
BSB 6289
Prediluted
Ready-To-Use
3.0 ml
BSB 6290
Prediluted
Ready-To-Use
7.0 ml
BSB 6291
Prediluted
Ready-To-Use
15.0 ml
BSB 6292
Concentrated
1:500-1:2000
0.1 ml
BSB 6293
Concentrated
1:500-1:2000
0.5 ml
BSB 6294
Concentrated
1:500-1:2000
1.0 ml
BSB 6295
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Liang Y, Johansson O, J Invest Dermatol. 1998;Jul;111(1):114-8
  2. Radfar A, et al. Am J Dermatopathol. 2006;Apr;28(2):162-7

Recommended Immunohistochemical Protocol

Pretreatment
  1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
  2. Air dry for 2 hours at 58° C.
  3. Deparaffinize, dehydrate and rehydrate tissues.
  4. Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
  5. Any of three heating methods may be used:
    • Electric Pressure Cooker
      Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature.
    • Water Bath Method
      Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes.
    • Conventional Steamer Method
      Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
  6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
  7. Wash slides with IHC wash buffer or DI water.
  8. Continue IHC staining protocol.

Immunohistochemical Protocol

Step

ImmunoDetector
(AP or HRP)

PolyDetector
(AP or HRP)
Peroxidase/AP Block
5 minutes
5 minutes
Primary Antibody
30 minutes
45 minutes
Secondary Biotinylated Link
10 minutes
Not Applicable
AP or HRP Label
10 minutes
45 minutes
Substrate-Chromogen
5-10 minutes
10 minutes
Counterstaining
Time varies with counterstain
Time varies with counterstain