QuantiMatrix® Human Laminin ELISA Kit 84 Determinations

Analytes Available	Species Reactivity	Key Applications	Detection Methods
Laminin	H	ELISA	Chromogenic

Analytes Available

Species Reactivity

Key Applications

Detection Methods

Laminin

ELISA

Chromogenic

Description
Catalogue Number	ECM310
Brand Family	Chemicon®
Trade Name	Quantimatrix Chemicon
Description	QuantiMatrix Human Laminin ELISA
Overview	The QuantiMatrix™ ELISA kit for the determination of soluble human laminin provides sensitivity down to 30-70 ng/mL as determined from culture supernatants and purified protein samples. The assay is in the format of a competitive inhibition ELISA. Human laminin is pre-coated on the wells with the exception of row H, which serves as a reference. Standards and samples are diluted and pre-incubated with polyclonal rabbit antibody to human laminin. The polyclonal antibody binds to laminin in the standards and samples. The mixture is then transferred to the human laminin-coated plate. Free rabbit anti-human laminin binds to the laminin on the plate. Goat anti-rabbit IgG HRP conjugate reacts with bound rabbit anti-laminin. When HRP substrate is added, a blue color develops. The reaction is stopped by the addition of an acid, changing the color to yellow. This color is quantitated using an ELISA reader; the intensity of the color is inversely proportional to the concentration of laminin in the original sample. A standard curve is constructed and sample values are interpolated. ECM310 QuantiMatrix Human Laminin ELISA kit is prepared with antibody made against pepsinized human placental laminin and the Control Standard in the kit is also a pepsinized placental laminin preparation. The most common laminin chains expressed in human placenta are generally the alpha 1, alpha 2 and beta 1, beta 2 chains of Laminin. Laminin 5 (alpha3beta3gamma2) is also present in placenta but we have not tested the ECM310 specifically with Laminin 5. It is possible that the kit would detect laminin 5 but we suspect that the signal for laminin 5 might be very weak. The strongest signals would probably be expressed with laminins 1 through 4.
Background Information	The determination of levels of extracellular matrix molecules is becoming more important to understanding alterations in matrix function during development and tumorigenesis. Questions of matrix gene regulation and function require a sensitive means of determining the levels of protein expression by cells.

Description

Catalogue Number

ECM310

Brand Family

Chemicon®

Trade Name

Quantimatrix
Chemicon

Description

QuantiMatrix Human Laminin ELISA

Overview

The QuantiMatrix™ ELISA kit for the determination of soluble human laminin provides sensitivity down to 30-70 ng/mL as determined from culture supernatants and purified protein samples.

The assay is in the format of a competitive inhibition ELISA. Human laminin is pre-coated on the wells with the exception of row H, which serves as a reference. Standards and samples are diluted and pre-incubated with polyclonal rabbit antibody to human laminin. The polyclonal antibody binds to laminin in the standards and samples. The mixture is then transferred to the human laminin-coated plate. Free rabbit anti-human laminin binds to the laminin on the plate. Goat anti-rabbit IgG HRP conjugate reacts with bound rabbit anti-laminin. When HRP substrate is added, a blue color develops. The reaction is stopped by the addition of an acid, changing the color to yellow. This color is quantitated using an ELISA reader; the intensity of the color is inversely proportional to the concentration of laminin in the original sample. A standard curve is constructed and sample values are interpolated.

ECM310 QuantiMatrix Human Laminin ELISA kit is prepared with antibody made against pepsinized human placental laminin and the Control Standard in the kit is also a pepsinized placental laminin preparation. The most common laminin chains expressed in human placenta are generally the alpha 1, alpha 2 and beta 1, beta 2 chains of Laminin. Laminin 5 (alpha3beta3gamma2) is also present in placenta but we have not tested the ECM310 specifically with Laminin 5. It is possible that the kit would detect laminin 5 but we suspect that the signal for laminin 5 might be very weak. The strongest signals would probably be expressed with laminins 1 through 4.

Background Information

The determination of levels of extracellular matrix molecules is becoming more important to understanding alterations in matrix function during development and tumorigenesis. Questions of matrix gene regulation and function require a sensitive means of determining the levels of protein expression by cells.

Product Information
Components	QuantiMatrixTM Laminin Coated Strips (Part No. 60116) - 1 x 96-well strip plate Human Laminin Standard (Part No. 60117) - 1 lyophilized bottle; reconstitute as indicated on label Sample Diluent (Part No. 60095) - 1 x 15 mL bottle of 1% BSA in PBS + 0.01% Thimerosal Goat anti-Rabbit IgG-HRP Conjugate (QM2) (Part No. CS216103) - 1 bottle; ready to use Rabbit anti-Human Laminin (Part No. 60118) - 1 bottle polyclonal antibody; ready to use TMB/E Substrate (Part No. 60096) - 1 bottle; ready to use Stop Solution (Part No. 60193) - 1 bottle; (0.5M HCl solution) Wash Buffer Concentrate (Part No. 60245) - 1 x 100 mL bottle; provides final concentrations of 0.05% Tween-20 and 0.01% Thimerosal when added to 1 L of deionized water as described in step 1 of protocol on page 2.
Detection method	Chromogenic

Product Information

Components

QuantiMatrixTM Laminin Coated Strips (Part No. 60116) - 1 x 96-well strip plate
Human Laminin Standard (Part No. 60117) - 1 lyophilized bottle; reconstitute as indicated on label
Sample Diluent (Part No. 60095) - 1 x 15 mL bottle of 1% BSA in PBS + 0.01% Thimerosal
Goat anti-Rabbit IgG-HRP Conjugate (QM2) (Part No. CS216103) - 1 bottle; ready to use
Rabbit anti-Human Laminin (Part No. 60118) - 1 bottle polyclonal antibody; ready to use
TMB/E Substrate (Part No. 60096) - 1 bottle; ready to use
Stop Solution (Part No. 60193) - 1 bottle; (0.5M HCl solution)
Wash Buffer Concentrate (Part No. 60245) - 1 x 100 mL bottle; provides final concentrations of 0.05% Tween-20 and 0.01% Thimerosal when added to 1 L of deionized water as described in step 1 of protocol on page 2.

Detection method

Chromogenic

Applications
Key Applications	ELISA

Applications

Key Applications

ELISA

Biological Information
Species Reactivity	Human
Analytes Available	Laminin
Entrez Gene Number	NM_002291.1
Entrez Gene Summary	Laminins, a family of extracellular matrix glycoproteins, are the major noncollagenous constituent of basement membranes. They have been implicated in a wide variety of biological processes including cell adhesion, differentiation, migration, signaling, neurite outgrowth and metastasis. Laminins are composed of 3 non identical chains: laminin alpha, beta and gamma (formerly A, B1, and B2, respectively) and they form a cruciform structure consisting of 3 short arms, each formed by a different chain, and a long arm composed of all 3 chains. Each laminin chain is a multidomain protein encoded by a distinct gene. Several isoforms of each chain have been described. Different alpha, beta and gamma chain isomers combine to give rise to different heterotrimeric laminin isoforms which are designated by Arabic numerals in the order of their discovery, i.e. alpha1beta1gamma1 heterotrimer is laminin 1. The biological functions of the different chains and trimer molecules are largely unknown, but some of the chains have been shown to differ with respect to their tissue distribution, presumably reflecting diverse functions in vivo. This gene encodes the beta chain isoform laminin, beta 1. The beta 1 chain has 7 structurally distinct domains which it shares with other beta chain isomers. The C-terminal helical region containing domains I and II are separated by domain alpha, domains III and V contain several EGF-like repeats, and domains IV and VI have a globular conformation. Laminin, beta 1 is expressed in most tissues that produce basement membranes, and is one of the 3 chains constituting laminin 1, the first laminin isolated from Engelbreth-Holm-Swarm (EHS) tumor. A sequence in the beta 1 chain that is involved in cell attachment, chemotaxis, and binding to the laminin receptor was identified and shown to have the capacity to inhibit metastasis.
Gene Symbol	LAMB1 MGC142015 CLM
UniProt Number	P07942
UniProt Summary	FUNCTION: SwissProt: P07942 # 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. SIZE: 1786 amino acids; 198066 Da 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. Beta-1 is a subunit of laminin-1 (EHS laminin), laminin-2 (merosin), and laminin-6 (K-laminin). SUBCELLULAR LOCATION: Secreted, extracellular space, extracellular matrix, basement membrane. Note=Major component. DOMAIN: SwissProt: P07942 The alpha-helical domains I and II are thought to interact with other laminin chains to form a coiled coil structure. & Domains VI and IV are globular. SIMILARITY: Contains 13 laminin EGF-like domains. & Contains 1 laminin IV type B domain. & Contains 1 laminin N-terminal domain.

Biological Information

Species Reactivity

Human

Analytes Available

Laminin

Entrez Gene Number

NM_002291.1

Entrez Gene Summary

Laminins, a family of extracellular matrix glycoproteins, are the major noncollagenous constituent of basement membranes. They have been implicated in a wide variety of biological processes including cell adhesion, differentiation, migration, signaling, neurite outgrowth and metastasis. Laminins are composed of 3 non identical chains: laminin alpha, beta and gamma (formerly A, B1, and B2, respectively) and they form a cruciform structure consisting of 3 short arms, each formed by a different chain, and a long arm composed of all 3 chains. Each laminin chain is a multidomain protein encoded by a distinct gene. Several isoforms of each chain have been described. Different alpha, beta and gamma chain isomers combine to give rise to different heterotrimeric laminin isoforms which are designated by Arabic numerals in the order of their discovery, i.e. alpha1beta1gamma1 heterotrimer is laminin 1. The biological functions of the different chains and trimer molecules are largely unknown, but some of the chains have been shown to differ with respect to their tissue distribution, presumably reflecting diverse functions in vivo. This gene encodes the beta chain isoform laminin, beta 1. The beta 1 chain has 7 structurally distinct domains which it shares with other beta chain isomers. The C-terminal helical region containing domains I and II are separated by domain alpha, domains III and V contain several EGF-like repeats, and domains IV and VI have a globular conformation. Laminin, beta 1 is expressed in most tissues that produce basement membranes, and is one of the 3 chains constituting laminin 1, the first laminin isolated from Engelbreth-Holm-Swarm (EHS) tumor. A sequence in the beta 1 chain that is involved in cell attachment, chemotaxis, and binding to the laminin receptor was identified and shown to have the capacity to inhibit metastasis.

Gene Symbol

LAMB1
MGC142015
CLM

UniProt Number

P07942

UniProt Summary

FUNCTION: SwissProt: P07942 # 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.
SIZE: 1786 amino acids; 198066 Da
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. Beta-1 is a subunit of laminin-1 (EHS laminin), laminin-2 (merosin), and laminin-6 (K-laminin).
SUBCELLULAR LOCATION: Secreted, extracellular space, extracellular matrix, basement membrane. Note=Major component.
DOMAIN: SwissProt: P07942 The alpha-helical domains I and II are thought to interact with other laminin chains to form a coiled coil structure. & Domains VI and IV are globular.
SIMILARITY: Contains 13 laminin EGF-like domains. & Contains 1 laminin IV type B domain. & Contains 1 laminin N-terminal domain.

Product Usage Statements
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Store the kit and all components at 2-8°C. PRECAUTIONS: Human Laminin was purified from human tissue from donors tested negative for HBsAg, HIV and antibody to HIV. However, all samples and materials should be handled carefully and in accordance with good laboratory practices. The instructions provided have been designed to optimize performance. Deviations from the suggested procedure may result in sub-optimal performance of the kit and failure to produce accurate data. Do not mix or substitute reagents from different kit lots.

Storage and Shipping Information

Storage Conditions

Store the kit and all components at 2-8°C.

PRECAUTIONS: Human Laminin was purified from human tissue from donors tested negative for HBsAg, HIV and antibody to HIV. However, all samples and materials should be handled carefully and in accordance with good laboratory practices.

The instructions provided have been designed to optimize performance. Deviations from the suggested procedure may result in sub-optimal performance of the kit and failure to produce accurate data.

Do not mix or substitute reagents from different kit lots.

Packaging Information
Material Size	1 kit

Packaging Information

Material Size

1 kit

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

QuantiMatrix® Human Laminin ELISA Kit 84 Determinations

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