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Actin, Muscle-Specific

IHC of Actin, Muscle-Specific on an FFPE Skeletal Muscle Tissue

Description

Actin is a globular-structural, 345 kDa protein that polymerizes in a helical fashion to form an actin filament (or microfilament). Actin filaments provide mechanical support for the cell, determine the cell shape, enable cell movements (through lamellipodia, filopodia, or pseudopodia); and participate in certain cell junctions, in cytoplasmic streaming and in contraction of the cell during cytokinesis. In muscle cells they play an essential role, along with myosin, in muscle contraction. In the cytosol, actin is predominantly bound to ATP, but can also bind to ADP.


This antibody recognizes actin of skeletal, cardiac, and smooth-muscle cells. It is not reactive with other mesenchymal cells except for myoepithelium. Muscle-Specific Actin recognizes alpha and gamma isotypes of all muscle groups. Non-muscle cells such as vascular endothelial cells and connective tissues are nonreactive. Neoplastic cells of non-muscle-derived tissue such as Carcinomas, Melanomas and Lymphomas are negative. This antibody is useful in the identification of rhabdoid cellular elements.

Antibody Type
Mouse Monoclonal
Clone
HHF-35
Isotype
IgG1/K
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Skeletal Muscle
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

Actin, Muscle Specific 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 5022
Prediluted
Ready-To-Use
3.0 ml
BSB 5023
Prediluted
Ready-To-Use
7.0 ml
BSB 5024
Prediluted
Ready-To-Use
15.0 ml
BSB 5025
Concentrated
1:25-1:100
0.1 ml
BSB 5026
Concentrated
1:25-1:100
0.5 ml
BSB 5027
Concentrated
1:25-1:100
1.0 ml
BSB 5028
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Gown, et al. A. J. P. 1986;125:191
  2. Schmidt R., et al. A. J. P. 1988;131:199
  3. Azumi N, et al. Modern Pathology. 1988,1:469-474
  4. Rangdaeng L, et al. Am J Clin Pathology. 1991;96:32-45
  5. Schmidt R, Cone R, Haas J, Gown A, Amer J Pathol. 1988;131:19

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