Human Anti-Human Placenta Type V Collagen IgG Antibody Assay Kit, TMB

Human Anti-Human Placenta Type V Collagen IgG Antibody Assay Kit, TMB - Cat Number: 20851T From Chondrex.

Research Field: Arthritis, Immunology

Clonality: N/A

Cross-Reactivity:

Host Origin: N/A

Applications: N/A

Isotype: N/A

Detection Range: 16 units/ml-0.25 units/ml

Sample Type: Serum, Plasma

Concentration: N/A

Immunogen:

PRODUCT SPECIFICATIONS

DESCRIPTION: Assay kit to quantify human/monkey anti-collagen antibodies
FORMAT: Pre-coated 96-well ELISA Plate with removeable strips
ASSAY TYPE: Indirect ELISA
ASSAY TIME: 4.5 hours
STANDARD RANGE: 16 units/ml to 0.25 units/ml
NUMBER OF SAMPLES: Up to 38 (duplicate) samples/standard plate (will vary for custom kits)
SAMPLE TYPES: Serum and Plasma
RECOMMENDED SAMPLE DILUTIONS: 1:100 (at least)
CHROMOGEN: OPD (read at 490 nm)
STORAGE: -20°C
VALIDATION DATA: N/A
NOTES: This kit has an overnight incubation step

INTRODUCTION

Human serum, especially from patients with autoimmune diseases, contain high levels of immunoreactive components, which yield high background levels in ELISA systems. These non-specific reactions are caused by adhesive immunoglobulins in human serum which so strongly adhere to plastic surfaces by hydrophilic binding that blocking agents such as bovine serum albumin (BSA) and Tween 20 are not capable of blocking these non-specific reactions at all. False positive reactions caused by the serum samples themselves are usually overlooked and are considered real antibody-antigen reactions in many cases, even now. In order to obtain the real values of antigenantibody reactions, it is critical 1) to choose proper blocking agents which block these kinds of non-specific reactions effectively, 2) to determine the unique non-specific background value of each individual sample using antigen-non-coated wells, and 3) to subtract that background value from the corresponding value determined in antigen-coated wells. In addition, it is important to determine the non-specific reactions caused by the secondary antibody as well. Chondrex, Inc.’s ELISA system incorporates unique blocking agents that inhibit the hydrophobic binding of these serum components onto plastic surfaces and are designed to determine the background values of individual samples using antigen-non-coated wells. Type V collagen is a minor collagen component that forms interstitial collagen fibrils with type I collagen and plays a critical role in the regulation of fibrillogenesis. Type V collagen from most tissues consists of two α1(V) chains and one α2(V) chain, [α1(V)]2α2(V), whereas the placenta contains two subtypes of type V collagen, [α1(V)]2α2(V) and α1(V)α2(V)α3(V). Immune reactions to type V collagen may play a role in graft rejection following lung transplant. For studying the possible involvement of type V collagen immunity in organ allograft rejection in both in vitro and in vivo experiments, Chondrex Inc. provides type V collagen-coated ELISA plates (wells) in addition to uncoated wells for use as blanks. This ELISA kit contains enough materials to run two plates on two separate occasions and may be used for monkey serum as well as human serum.

KIT COMPONENTS

PLATE COATING AND SETUP

Standard ELISA Kit with One Species of Type I or Type II Collagen

A standard ELISA kit consisting of five 8-well strips which are uncoated and serve as a control for background

levels of individual samples, five 8-well strips coated with one species of type V collagen to determine specific antibody levels,

and two 8- well strips for reference standards. “B” represents blank wells to determine non-specific reactions caused by the

secondary antibody. Standards and samples are run in duplicate.

ASSAY OUTLINE