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CD56

IHC of CD56 on an FFPE Neuroblastoma Tissue

Description

CD56 or Neural-Cell Adhesion Molecule (NCAM) is a homophilic binding glycoprotein expressed on the surface of neurons, glia and skeletal muscle. CD56 has been implicated in cell-cell adhesion, neurite outgrowth, synaptic plasticity, and learning and memory.


Normal cells that stain positively for CD56 include NK cells, activated T-cells, brain and cerebellum, and neuroendocrine tissues. Tumors that are CD56-positive are Myeloma, Myeloid Leukemia, Neuroendocrine tumors, Wilm’s Tumor, Adult Neuroblastoma, NK/T cell Lymphomas, Pancreatic Acinar-cell Carcinoma, Pheochromocytoma, and Small-cell Lung Carcinoma. It is also expressed on some mesodermally-derived tumors (Rhabdomyosarcoma). Ewing’s Sarcoma/PNET is CD56-negative.

Antibody Type
Mouse Monoclonal
Clone
123C3.D5
Isotype
IgG1/K
Reactivity
Paraffin, Frozen
Localization
Membranous
Control
Neuroblastoma
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

CD56 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 5267
Prediluted
Ready-To-Use
3.0 ml
BSB 5268
Prediluted
Ready-To-Use
7.0 ml
BSB 5269
Prediluted
Ready-To-Use
15.0 ml
BSB 5270
Concentrated
1:200-1:800
0.1 ml
BSB 5271
Concentrated
1:200-1:800
0.5 ml
BSB 5272
Concentrated
1:200-1:800
1.0 ml
BSB 5273
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Gerardy-Schahn R, et al. International J of Cancer Sup. 1994;8:38-42
  2. Michalides R, et al. International J of Cancer Sup. 1994;8:34-37
  3. Kibbelaar RE, et al. Euro J of Cancer. 1991;27(4):431-435
  4. Moolenaar CE, et al. Cancer Research. 1990;50(4):1102-1106
  5. Langdon SP, et al. Cancer Research. 1988;48(21):6161-6165

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