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CD45

IHC of CD45 on an FFPE Tonsil Tissue

Description

The CD45 antigen is a protein which was originally called Leukocyte Common Antigen. It is a Type I transmembrane protein which is in various forms present on all differentiated hematopoietic cells except erythrocytes and assists in the activation of those cells (a form of co-stimulation). It is expressed in Lymphomas, B-cell Chronic Lymphocytic Leukemia, Hairy Cell Leukemia, and Acute Non-lymphocytic Leukemia.


CD45 is a monoclonal antibody that is routinely used to aid in the differential diagnosis of undifferentiated neoplasms, whenever malignant Lymphoma is suspected by the morphological or clinical data. It is a highly specific antibody; thus, a positive result is highly indicative of lymphoid or myeloid origin. Certain types of lymphoid neoplasms may lack CD45 (Hodgkin’s Disease, some T-cell Lymphomas and some Leukemias) so its absence does not rule out a hematolymphoid tumor. This antibody is exclusively expressed by cells of hematopoietic lineage and is present in most benign and malignant lymphocytes, erythrocytes and plasma cell precursors.

Antibody Type
Mouse Monoclonal
Clone
2B11 & PD7/26
Isotype
IgG1/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

CD45 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 5246
Prediluted
Ready-To-Use
3.0 ml
BSB 5247
Prediluted
Ready-To-Use
7.0 ml
BSB 5248
Prediluted
Ready-To-Use
15.0 ml
BSB 5249
Concentrated
1:500-1:2000
0.1 ml
BSB 5250
Concentrated
1:500-1:2000
0.5 ml
BSB 5251
Concentrated
1:500-1:2000
1.0 ml
BSB 5252
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Mason DY, Am Pathol. 1987;128:1-4
  2. Hall PA, Histopathology. 1988;13:149-160
  3. Kurtin PJ, Hum Path. 1985;16:353-365

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