PDF

CD61

IHC of CD61 on an FFPE Bone Marrow Tissue

Description

CD61 is a glycoprotein found on megakaryocytes (bone marrow cells), platelets and their precursors. CD61 antigen plays a role in platelet aggregation and also as a receptor for fibrinogen, fibronectin, von Willebrand factor and vitronectrin.


CD61 labels the IIIa subunit of the noncovalently-linked glycoprotein heterodimer IIb/IIIa complex present on human platelets and their precursors. This antibody is useful in identifying megakaryoblastic differentiation as seen in Megakaryoblastic Leukemia.

Antibody Type
Mouse Monoclonal
Clone
2f2
Isotype
IgG1/K
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Bone Marrow
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

Anti-CD61 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 5281
Prediluted
Ready-To-Use
3.0 ml
BSB 5282
Prediluted
Ready-To-Use
7.0 ml
BSB 5283
Prediluted
Ready-To-Use
15.0 ml
BSB 5284
Concentrated
1:50-1:200
0.1 ml
BSB 5285
Concentrated
1:50-1:200
0.5 ml
BSB 5286
Concentrated
1:50-1:200
1.0 ml
BSB 5287
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Thiele J, et al. Eur J Haematol. 1990;44:63-70
  2. Thiele J, et al. Virchows Archiv B Cell Pathol. 1990;58:295-302
  3. Goldman BI, et al. Modern Pathology. 2001;14:589-594
  4. Fox SB, et al. 1990;Jul; 17(1):69-74
  5. Duperray A, et al. Blood. 1989;Oct;74(5):1603-1611
  6. Campana D, et al. Leukemia. 1990;Sep;4(9):620-624
  7. Thiele J, et al. Anal Quant Histol. 1990;Aug;12(4):285-289
  8. Gatter KC, et al. Histopathology. 1998;13:257-267

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