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CD8

IHC of CD8 on an FFPE Tonsil Tissue

Description

CD8 is a transmembrane glycoprotein that serves as a co-receptor for the T-cell receptor (TCR). Like the TCR, CD8 binds to a major histocompatibility complex (MHC) molecule that is specific for the Class I MHC protein. To function, CD8 forms a dimer, consisting of a pair of CD8 chains. The most common form of CD8 is composed of a CD8-α and CD8-β, chain, both members of the immunoglobulin superfamily with an immunoglobulin variable (IgV)-like extracellular domain connected to the membrane by a thin stalk, and an intracellular tail.


CD8 is a T-cell marker for the detection of cytotoxic/suppressor cells of blood lymphocytes. CD8 is also detected on NK cells, most thymocytes, a subpopulation of null cells and bone marrow cells. This antibody is used to distinguish between reactive and neoplastic T-cells.

Antibody Type
Mouse Monoclonal
Clone
C8/144B
Isotype
IgG/K
Reactivity
Paraffin, Frozen
Localization
Membranous
Control
Tonsil, Lymph Node
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

CD8 is a mouse monoclonal antibody derived from tissue culture supernatant diluted in Phosphate Buffered Saline, pH 7.6, with protein base, and preserved with Sodium Azide preservative.

Availability
Catalog No.
Antibody Type
Dilution
Volume/QTY
BSB 5169
Prediluted
Ready-To-Use
3.0 ml
BSB 5170
Prediluted
Ready-To-Use
7.0 ml
BSB 5171
Prediluted
Ready-To-Use
15.0 ml
BSB 5172
Concentrated
1:250-1:1000
0.1 ml
BSB 5173
Concentrated
1:250-1:1000
0.5 ml
BSB 5174
Concentrated
1:250-1:1000
1.0 ml
BSB 5175
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Gao G, Jakobsen B, Immunol Today. 2000;21(12):630-636
  2. Rossi ML, Sanchez FC, et al. J Clin Path. 1988;41:314-319
  3. Stein H, Lennart K, et al. Adv Cancer Res. 1984;42:67-147
  4. Phan-Dinh-Tuy F, Niaudet P, et al. Mol Immun. 1982;19:1649-1654

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