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CD20

IHC of CD20 on an FFPE Tonsil Tissue

Description

CD20 is a transmembrane, non-glycosylated protein expressed on B-cell precursors and mature B-cells, but lost following differentiation into plasma cells. This antibody does not cross-react with non-hematopoietic neoplasms. CD20 (B-cell Pan) reacts with a membrane antigen present in B-cells.


This antibody strongly recognizes Reed-Sternberg cells predominant in Hodgkin’s disease. Since no staining of histiocytes or plasma cells has been observed and CD20 has not been detected in T-cell malignancies, it is a very strong marker of B-cell Lymphomas. B-cell Panmarker recognizes a formalin-resistant intracytoplasmic antigen.

Antibody Type
Mouse Monoclonal
Clone
L26
Isotype
IgG2a/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

CD20 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 5190
Prediluted
Ready-To-Use
3.0 ml
BSB 5191
Prediluted
Ready-To-Use
7.0 ml
BSB 5192
Prediluted
Ready-To-Use
15.0 ml
BSB 5193
Concentrated
1:500-1:2000
0.1 ml
BSB 5194
Concentrated
1:500-1:2000
0.5 ml
BSB 5195
Concentrated
1:500-1:2000
1.0 ml
BSB 5196
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Ishii Y, et al. Clin Exp Immuno. 1984;58:183-192
  2. Davey FR, et al. Am J Pathol. 1987;129:54-63
  3. Mason DY, Am J Pathol. 1987;128:1-4

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