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Fascin

IHC of Fascin on an FFPE Hodgkin’s Lymphoma Tissue

Description

Fascin, encoded by the human homolog for sn (hsn) gene, has been localized to microspikes and stress fibers of cultured cells where it is thought to be involved in the formation of microfilament bundles. It is expressed predominantly in dendritic cells. Lymphoid cells, myeloid cells and plasma cells are negative. However, Reed Sternberg cells in Hodgkin’s Lymphoma are positive for Fascin staining. Epstein-Barr virus may induce expression of Fascin in B-cells.


Fascin is a very sensitive marker for Reed-Sternberg cells and variants in nodular sclerosis, mixed cellularity, and lymphocyte depletion Hodgkin’s Disease. This marker might be helpful in distinguishing between Hodgkin’s Disease and Non-Hodgkin Lymphoma in difficult cases. Also, the lack of expression of Fascin in the neoplastic follicles in Follicular Lymphoma can be helpful in distinguishing these lymphomas from reactive Follicular Hyperplasia in which the number of follicular dendritic cells is normal or increased.

Antibody Type
Mouse Monoclonal
Clone
55-k2
Isotype
IgG1
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Hodgkin’s Lymphoma, Lymph Node, Tonsil
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

Fascin 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 5512
Prediluted
Ready-To-Use
3.0 ml
BSB 5513
Prediluted
Ready-To-Use
7.0 ml
BSB 5514
Prediluted
Ready-To-Use
15.0 ml
BSB 5515
Concentrated
1:50-1:200
0.1 ml
BSB 5516
Concentrated
1:50-1:200
0.5 ml
BSB 5517
Concentrated
1:50-1:200
1.0 ml
BSB 5518
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Pelosi G, et al. Lung Cancer. 2003;Nov;42(2):203213
  2. Goncharuk VN, et al. Cutan Pathol. 2002;Aug;29(7):430438

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