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CD23

IHC of CD23 on an FFPE Tonsil Tissue

Description

CD23, also known as Fc epsilon RII, is the “low affinity” receptor for IgE, an antibody isotype involved in allergy and (arguably) resistance to parasites, and is important in regulation of IgE levels. Unlike many of the antibody receptors, CD23 is a C-type lectin. It is found on mature B-cells, activated macrophages, eosinophils, follicular dendritic cells and platelets.


This is a B-cell antibody that is useful for differentiating between B-CLL and B-SLL’s that are CD23-positive from Mantle-cell Lymphomas and Small-Cleaved Lymphomas that are CD23- negative. This antibody reacts with the antigen that is found on a subpopulation of peripheral blood cells, B-lymphocytes and on EBV-transformed B-lymphoblastoid cell lines.

Antibody Type
Rabbit Monoclonal
Clone
SP23
Isotype
IgG
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

CD23 is a mouse monoclonal antibody 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 5204
Prediluted
Ready-To-Use
3.0 ml
BSB 5205
Prediluted
Ready-To-Use
7.0 ml
BSB 5206
Prediluted
Ready-To-Use
15.0 ml
BSB 5207
Concentrated
1:25-1:100
0.1 ml
BSB 5208
Concentrated
1:25-1:100
0.5 ml
BSB 5209
Concentrated
1:25-1:100
1.0 ml
BSB 5210
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Kaiserlian D, et al. Immunology. 1993;80:90-95
  2. Aubry JP, et al. Oxford Univ Press- Oxford, NY, Tokyo. 1987;417-419
  3. Pallesen G, Oxford Univ Press-Oxford, NY, Tokyo. 1987;383-386

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