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CD30

IHC of CD30 on an FFPE Hodgkin’s Lymphoma Tissue

Description

CD30 is a transmembrane cytokine receptor belonging to the tumor necrosis factor (TNF) receptor superfamily. Mature CD30 has a molecular mass of 120 kDa and is derived from a 90 kDa precursor protein.


CD30 antibody detects an epitope which is expressed by Reed-Sternberg cells in Hodgkin’s Disease, the majority of Anaplastic Large-cell Lymphomas, and in Embryonal Carcinomas and Seminomas. This antibody also stains plasma cells intensely in paraffin-embedded tissue.

Antibody Type
Mouse Monoclonal
Clone
Ber-H2
Isotype
IgG1/K
Reactivity
Paraffin, Frozen
Localization
Membranous
Control
Hodgkin’s Lymphoma
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

CD30 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 5211
Prediluted
Ready-To-Use
3.0 ml
BSB 5212
Prediluted
Ready-To-Use
7.0 ml
BSB 5213
Prediluted
Ready-To-Use
15.0 ml
BSB 5214
Concentrated
1:250-1:1000
0.1 ml
BSB 5215
Concentrated
1:250-1:1000
0.5 ml
BSB 5216
Concentrated
1:250-1:1000
1.0 ml
BSB 5217
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Schwarting R, et al. Blood. 1989;74:1678-1689
  2. Fonatsch C, et al. Genomics. 1992;14:825-826
  3. Piris J, et al. Histopathology. 1990;17:211-218

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