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Laminin EIA Kit



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Product Name: Laminin EIA Kit

# of Samples: 1X96 Assays

Intended Use: The Laminin EIA Kit is an in vitro enzyme immunoassay (EIA) kit for quantitative determination of human LN in plasma, serum, urine, cultured cell extracts, cell culture supernatants, and other biological fluids.
This kit is for research use only. It is not for use in diagnostic or therapeutic procedures.

Introduction: Laminins, which are large multidomain glycoproteins of the extracellular matrix, have attracted much attention because of their importance in cellular functions including induction of cell adhesion, growth promotion, mediation of cell communication, and enhancement of the metastatic phenotype of tumor cells (1). Laminins (LN) consist of three different polypeptide chains, ?,? and ? chain types, which replaced the previous designation of A, B1 and B2 (2). These chains are linked by disulfide bridges to form a characteristic asymmetric cross-structure as seen by electron microscopy (3). LN binds to various components of the basement membrane and probably links these to one another (4). Cell surface receptors that may play a role in LN-mediated cell adhesion have been isolated from metastatic tumor cells (5) and platelets(6). Most of the studies on LN have focused on the protein isolated from tumors (7), placenta (8), or products from cultured cell lines (9). Serum levels of LN fragments have been reported to be elevated in patients with hepatic fibrosis (10), alcholic liver (11), hypertension (12), and several kinds of tumors (13). Since LN was isolated also from urine and urinary LN was identified to be a fragmented ??1 chain (14) recently, the detection system having higher sensitivity has been required. The enzyme immunoassay system was developed for quantitative detection of LN by using domain-specific monoclonal antibodies. This kit uses ??1-specific monoclonal antibodies and can be used to detect quantitatively both serum and urinary LN. By usage of this kit, it has been shown that the levels of urinary LN fragments become elevated in some tumors (14).

Principle of the test: The Laminin EIA Kit is a solid phase EIA based on a sandwich method that utilizes two mouse monoclonal anti-LN antibodies to detect LN by two-step procedure. One of the antibodies is bound to a microtiter plate to create the solid phase. Non-specific binding is blocked by a blocking buffer. Samples and standards are incubated in microtiter-plate wells. After washing the plate, the second anti-LN which is labelled with peroxidase (POD) is added to the wells and incubated. During these steps, LN is captured onto the solid support on one side and tagged on the other by POD-anti-LN. The reaction between POD and substrate (H2O2, and tetramethylbenzidine) results in colour development with intensities proportional to the amount of LN present in samples and standards. The amount of LN can be quantitatively determined by measuring the absorbance using an EIA plate reader. Accurate sample concentrations of LN can be determined by comparing their specific absorbance with those obtained for the standards plotted on a standard curve.

Storage and Stability: This kit is shipped at 2 - 8C and should be stored at 2 - 8C if not used. Under this condition, the kit is stable until the expiration date on label.

Sensitivity and Specificity: This kit specifically measures human LN with no detectable cross reaction with human fibronectin, vitronectin, fibrinogen, collagen type I, or collagen type III. This kit can be also used to measure rabbit LN. The application of this kit for quantitative detection of LN from other sources has not been tested.

Product Name: Laminin EIA Kit

Function: Binding to cells via a high affinity receptor, laminin is thought to mediate the attachment, migration and organization of cells into tissues during embryonic development by interacting with other extracellular matrix components.

Subunit: Laminin is a complex glycoprotein, consisting of three different polypeptide chains (alpha, beta, gamma), which are bound to each other by disulfide bonds into a cross-shaped molecule comprising one long and three short arms with globules at each end. Alpha-1 is a subunit of laminin-1 (EHS laminin) and laminin-3 (S-laminin).

Subcellular Location: Secreted, extracellular space, extracellular matrix, basement membrane. Note=Major component.

Domain: The alpha-helical domains I and II are thought to interact with other laminin chains to form a coiled coil structure.

Domain: Domains VI, IV and G are globular.

Similarity: Contains 17 laminin EGF-like domains.

Similarity: Contains 5 laminin G-like domains.

Similarity: Contains 2 laminin IV type A domains.

Similarity: Contains 1 laminin N-terminal domain.

Additional Information

Name Laminin EIA Kit
Related Product Names Laminin EIA ELISA Kit; Laminin A chain LAMALaminin EIA KitLAMA1
NCBI Acc Number NP_005550.2
Molecular Weight 337158
Datasheets / Downloads GWB-783B25 Datasheet
Swiss Prot Number P25391
Intended Use Research Use Only