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. Porcine collagen type IV 100%, porcine collagens type I, II, and III <0.1%, porcine elastin, fibronectin and laminin < 0.1% (RIA at 1:5000 dilution).

Source
Rabbits were immunized with purified collagen type IV from porcine kidney tissue

Product
Affinity purified antibody lyophilized from phosphate buffered solution; no BSA and preservative added! Reconstitute with 0.1 ml sterile bidistilled water.

Secondary Reagents: Anti-rabbit IgG-conjugates, e.g. anti-rabbit IgG:FITC (Art. No. FI-1000) or anti-rabbit IgG:DyLight488 (Art. No. DI-1488).

Concentration: app. 1 mg/ml

Specificity

Species Reactivity: Pig, cross-reactivity with human < 0.1%, mouse and rat collagen IV <1%

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

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

Working Concentration: IFA 1:80, IHC(P) 1:500, ELISA >1:200

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: Porcine skin or liver

Storage
-20°C

Shipping Conditions: Ship at ambient temperature

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. Boucard N, Viton C, Agay D, Mari E, Roger T, Chancerelle Y, Domard A ( 2007) The use of physical hydrogels of chitosan for skin regeneration following third-degree burns. Biomaterials. Aug;28(24):3478-88.