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CD45RA

IHC of CD45RA on an FFPE Tonsil Tissue

Description

CD45 is a complex molecule and is comprised of different glycoproteins ranging from 180-240 kDa. Expression of CD45 is found on all hemopoietic cells. Detection of the different isoforms can distinguish between different cell forms (e.g., naive T-cells and memory T-cells). CD45RA is an isoform of the CD45 complex and has restricted expression between different subtypes of lymphoid cells.


CD45RA antibody reacts with mature, non-activated T and B-cells. CD45RA is also reactive with medullary thymocytes, mantle-zone lymphocytes in follicles of lymph nodes, spleen and lymphocytes of the paracortex. CD45RA shows no reactivity with cortical thymocytes, immature T-cells or activated B-cells in germinal centers.

Antibody Type
Mouse Monoclonal
Clone
111-1C5
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

CD45RA 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 5253
Prediluted
Ready-To-Use
3.0 ml
BSB 5254
Prediluted
Ready-To-Use
7.0 ml
BSB 5255
Prediluted
Ready-To-Use
15.0 ml
BSB 5256
Concentrated
1:25-1:100
0.1 ml
BSB 5257
Concentrated
1:25-1:100
0.5 ml
BSB 5258
Concentrated
1:25-1:100
1.0 ml
BSB 5259
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. N/A

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