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CD35

IHC of CD35 on an FFPE Tonsil Tissue

Description

CD35 (erythrocyte complement receptor 1 or CR1, also known as C3b/C4b receptor and immune adherence receptor) serves as the main system for processing and clearance of complement- opsonized immune complexes. The number of CR1 molecules decreases with aging of erythrocytes in normal individuals and is also decreased in pathological conditions such as Systemic Lupus Erythematosus (SLE), HIV infection, some Hemolytic Anemias and other conditions featuring immune complexes.


Anti-CD35 is considered a mature B-cell marker, which labels follicular dendritic reticulum cells and tumors derived from such cells such as Follicular Dendritic Cell Tumor/Sarcoma. CD35 antigen is found in erythrocytes, B-cells, and a subset of T-cells, monocytes, as well as in eosinophils and neutrophils.

Antibody Type
Mouse Monoclonal
Clone
RLB25
Isotype
IgG2b
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

Anti-CD35 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 5232
Prediluted
Ready-To-Use
3.0 ml
BSB 5233
Prediluted
Ready-To-Use
7.0 ml
BSB 5234
Prediluted
Ready-To-Use
15.0 ml
BSB 5235
Concentrated
1:25-1:100
0.1 ml
BSB 5236
Concentrated
1:25-1:100
0.5 ml
BSB 5237
Concentrated
1:25-1:100
1.0 ml
BSB 5238
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Dillon KM, et al. J Clin Pathol. 2002;Oct;55(10):791-4
  2. Pileri SA, et al. Histopathology. 2002;41;1-29
  3. Kunihiko M, et al. J Histochem Cytochem. 2002;50:1475-1485
  4. Chan AC, et al. Histopathology. 2001;Jun;38(6):510-8
  5. Biddle DA, et al. Modern Pathology. 2002;15:50-58
  6. Cheuk W, et al. Am J Surg Pathol. 2001;Jun;25(6):721-31
  7. Chang KC, et al. J Pathol. 2003;Nov;201(3):404-12

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