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CD25

IHC of CD25 on an FFPE Tonsil Tissue

Description

CD25 is the alpha chain of the IL-2 receptor. It is a Type I transmembrane protein present on activated T-cells, activated B-cells, some thymocytes, myeloid precursors, and oligodendrocytes that associates with CD122 to form a heterodimer that can act as a high-affinity receptor for IL-2. It is expressed in most B-cell neoplasms, some Acute Non-lymphocytic Leukemias, and Neuroblastomas.


Expression of CD25 is a reliable diagnostic tool for distinguishing neoplastic mast-cell aggregates from reactive proliferations, and has, therefore, recently become a minor criterion for the diagnosis of Systemic Mastocytosis (SM). Anti-CD25 antibodies have also been useful in identifying mast cells in skin biopsies in the setting of Urticaria Pigmentosa, which is predictive of Systemic Mastocytosis. Quantitation of regulatory T-cells (Treg) in the setting of hepatocellular carcinoma has been used as an independent predictive factor for tumor recurrence after hepatic resection for HCC. Also, the percentage of tumor-infiltrating CD25+FOXP3+ regulatory T-cells among tumor cells, inside tumor parenchyma and at its periphery are significantly higher in recurrent Cutaneous Melanoma than in Non-recurrent Melanoma.

Antibody Type
Mouse Monoclonal
Clone
4C9
Isotype
IgG2b
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic, Membranous
Control
Mastocytosis, Tonsil, Small Bowel
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

CD25 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 6317
Prediluted
Ready-To-Use
3.0 ml
BSB 6318
Prediluted
Ready-To-Use
7.0 ml
BSB 6319
Prediluted
Ready-To-Use
15.0 ml
BSB 6320
Concentrated
1:25-1:100
0.1 ml
BSB 6321
Concentrated
1:25-1:100
0.5 ml
BSB 6322
Concentrated
1:25-1:100
1.0 ml
BSB 6323
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Hahn HP, Hornick JL, Am J Surg Pathol. 2007;Nov;31(11):1669-76
  2. Hollmann TJ, et al. Am J Surg Pathol. 2008;Jan;32(1):139-45
  3. Miracco C, et al. Oncol Rep. 2007;Nov;8(5):1115-22
  4. Siddiqui SA, at al. Clin Cancer Res. 2007;Apr1;13(7):2075-81

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