Catalogue

Background
Type IV Collagen is a non-fibrillary network of different alpha-chains: alpha 1(IV) to-alpha 6(IV). It is typically found in basal membranes of different organs (e.g. skin, lens, lung, kidney). Collagens consist of a family of highly specialized glycoproteins of which at least 16 genetically distinct types are known to date. The basal unit of a collagen molecule consists of a triple-helical structure formed by 3 alpha-chains. Predominant amino acids are glycine, proline and hydroxproline. Regularly also lysines and hydroxylysines occur, which are responsible for cross-linkage and glycosylation of the protein chains. Different composition of alpha-chains and different glycosylation contribute to the high variability of collagens in different tissues and organs. Human collagen type IV 100%, human collagen types I, II, III, and V <0.1%, human fibronectin and laminin < 0.1% (RIA at 1:10000 dilution).

Source
Mouse monoclonal antibody obtained from fusion of splenocytes of animals immunized with purified collagen type IV from human placenta.

Immunogen: Purified collagen type IV from human placenta

Product
0.1 ml affinity purified antibody prod. in nude mice; lyophilized from phosphate buffered solution; no BSA and preservative added! Reconstitute in 0.1 ml sterile, distilled water.

Purification Method: 0.1 ml ProtG affinity purified antibody prod. in nude mice; lyophilized from phosphate buffered solution; no BSA and preservative added!

Secondary Reagents: Anti-mouse IgG-conjugates, e.g. Anti-mouse IgG:FITC (Art. No. FI-2000) or anti-mouse IgG:DyLight488 (Art. No. DI-2488).

Concentration: app. 0.2-0.4 mg/ml

Specificity

Species Reactivity: Human, no cross-reactivity with bovine, mouse, rat and porcine collagens type IV

Applications
IHC(P), IFA, ELISA, RIA, IB/WB

Incubation Time: IHC(P) 60 min at RT or 2-8°C over night

Working Concentration: (purified, lyophilized) IFA ? 1:320, IHC(P) ? 1:4000, ELISA ? 1:4000 (OD ? 500)

Pre-Treatment: After de-waxing the tissue slices they are treated with 0.2% hyaluronidase (app. 300 U/mg e.g. Art. No. HYA02-50) in TBS 15 min at 37°C. Thereafter non-specific binding is blocked by blocking serum or 3% BSA in TBS. For peroxidase systems blocking with 1% peroxide solution in TBS for 30 min at RT is recommended.

Positive Control: Human skin, placenta or lens capsule

Storage
-20°C

Caution
This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but Exalpha Biologicals accepts no liability for any inaccuracies or omissions in this information.

References
1. Guerret S, Govignon E, Hartmann DJ, Ronfard V. (2003) Long-term remodeling of a bilayered living human skin equivalent (Apligraf) grafted onto nude mice: immunolocalization of human cells and characterization of extracellular matrix. Wound Repair Regen. Jan-Feb;11(1):35-45 2. Rousseau C., Thillou F., Pagnon-Minot A., Guerret S., Hartmann D.J, Ronfard V. (2009) Short term integration on human bilayered living skin substitute (Apligraf®) grafted onto nude mice - 19th Annual Meeting of the Wound Healing Society with SAWC (Symposium on Advanced Wound Care) 2009 April 26-29, Gaylord Texan Resort and Convention Center, Dallas, Texas.

Protein Reference(s)

Database Name: UniProt

Accession Number: P02462 (CO4A1_HUMAN) ; P08572 (CO4A2_HUMAN); P2940

Species Accession: Human