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Collagen Type IV

IHC of Collagen IV on an FFPE Skin Tissue

Description

Collagen is the main protein of connective tissue in animals and the most abundant protein in mammals, making up about 25% of the total protein content. Collagen IV is a major constituent of the basement membranes, along with laminins and enactins. It is composed of the alpha 1 IV chain and alpha 2 IV chain in a 2:1 ratio. It can form insoluble fibers with high tensile strength.


Normal tissue stains with this antibody in a manner consistent with the sites of mesenchymal elements and epithelial basal laminae. Antibody to collagen IV is useful in detecting the loss of parts of basement membrane in carcinomas. Collagen IV can also be useful in the classification of soft tissue tumors; Schwanomas, Leiomyomas, and their well-differentiated malignant counterparts usually immunoreact to this antibody. The vascular nature of neoplasms, Hemangiopericytoma, Angiosarcoma and Epithelioid Hemangioendothelioma can be observed with this antibody.

Antibody Type
Mouse Monoclonal
Clone
CIV22
Isotype
IgG1/K
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Muscle, Lung
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

Collagen Type IV 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 5351
Prediluted
Ready-To-Use
3.0 ml
BSB 5352
Prediluted
Ready-To-Use
7.0 ml
BSB 5353
Prediluted
Ready-To-Use
15.0 ml
BSB 5354
Concentrated
1:10-1:30
0.1 ml
BSB 5355
Concentrated
1:10-1:30
0.5 ml
BSB 5356
Concentrated
1:10-1:30
1.0 ml
BSB 5357
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Gould VE, et al. Pathol Annual. 1976;11:353-386
  2. McArdle JP, et al. Int J Cancer. 1984;34:633-638
  3. Sakr WA, et al. Hum Path. 1987;18:1043-1050
  4. Barsky SH, et al. Am J Surg Pathol. 1983;7:667-677

Recommended Immunohistochemical Protocol

Pretreatment
with Enzyme
Digestion
  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 enzymatic digestion with ImmunoDNA Digestor (BSB 0108 – BSB 0112) or similar product (Proteinase K) using the following protocol:
    • Pre-warm the ImmunoDNA Digestor solution to room temperature
    • Incubate tissue sections with the ImmunoDNA Digestor solution for 10 minutes at room temperature or for 5 minutes at 37° C.
NOTE: 
Other enzyme treatments may reduce or eliminate the intensity of this antibody.
Heat induced epitope retrieval (HIER) yields poor results with this antibody.
  1. Wash slides with IHC wash buffer or DI water.
  2. 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