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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | ELISA, IH(P), WB | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB19562 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | Anti-Laminin-5 (γ2 chain) Antibody, clone D4B5 |
| Alternate Names |
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| Background Information | Laminins are basement membrane, extracellular matrix glycoproteins that are composed of alpha, beta and gamma heterotrimeric chains. Different combinations of five alpha chains, three beta chains, and two gamma chains form at least 12 laminin isoforms. Laminin-5 (LN5), which consists of laminin alpha3, beta3 and gamma2 chains, is a highly conserved component of anchoring filaments underlying the hemidesmosome structure of epidermal keratinocytes. Mutations or deletion in the LN5 genes (LAMA3, LAMB3, and LAMC2) is associated with epidermolysis bullosa, a lethal skin blistering disease. LN5 may stabilize the epidermal/dermal junction of skin through binding with integrins alpha3/beta1, alpha6/beta4 and type VII collagen and has a critical role in the anchorage and locomotion of a wide variety of cell types, including epithelial cells, fibroblasts, neurons and leukocytes. Compared to fibronectin, collagen, or vitronectin, cells of epithelial origin will adhere to Laminin-5 faster and will spread to a larger extent. Recently, Laminin-5 is attracting attention as a substrate for carcinoma cells that may stimulate migration, since it has been detected at the leading edge of invasive cancer tissue in several types of human carcinomas (Niki, et al., 2002, Lohi, et al., 2001). |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Protein A Purified mouse immunoglobulin in 20 mM sodium phosphate, 250 mM NaCl, pH. 7.6, with 0.1% sodium azide as a preservative. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Laminin-5 (γ2 chain) Antibody, clone D4B5 is an antibody against Laminin-5 (γ2 chain) for use in ELISA, IH(P) & WB. |
| Key Applications |
|
| Application Notes | Western blot: 5ug/ml is recommended; dilute antibody in PBS or TBS only, not blocking solutions; blocking with purified compounds is suggested. Immunohistochemistry of paraffin sections: 5-10ug/ml is suggested. Do not over fix. A light 4% or 2% PFA are best. 4 mm thick paraffin sections are dewaxed, rehydrated and immersed in 0.3% hydrogen peroxide-containing methanol for inactivation of intrinsic peroxidase, followed by treatment with Protease XXIV (Sigma) for 15 min at room temperature (0.01-0.05%). Do not use trypsin. Antibody incubation at 37°C for 1 hour, followed by secondary antibody detection and visualization with DAB (Mizushima et al. 1998). Do not place the monoclonal antibody in serum blocks because serum conatins laminin 5 which will bind and interfere with the antibody. Use BSA or other such media instead. Strongly positive staining is seen in Basement Membrane of the following tissues: Skin, Esophagus, Lung, and Kidney. Positive staining is also seen in Stomach, Small intestine, Spleen, Thymus, Prostate, and Ovary. Thyroid, Testis, Skeletal and Cardiac muscle were negative for LN5 staining. Inhibition of alpha2beta1 integrin binding to laminin 5 (40ug/ml) (Decline & Rousselle 2000). ELISA Optimal working dilutions must be determined by end user. |
| Biological Information | |
|---|---|
| Immunogen | Amino acid residues 382-608 of human laminin-5 gamma2 chain (III domain) expressed as a GST fusion protein |
| Clone | D4B5 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | Monoclonal antibody MAB19562 was developed against amino acid residues 382-608 of human laminin-5 gamma2 chain (III domain) expressed as a GST fusion protein. The antibody recognizes the 150 (precursor form) and 105 kDa (mature form) chain proteins of Laminin-5. No reactivity is expected with other laminin types, as the gamma2 chain of laminin is unique to laminin-5. |
| Isotype | IgG1 |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein A Purfied |
| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: Q13753 # 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. Ladsin exerts cell- scattering activity toward a wide variety of cells, including epithelial, endothelial, and fibroblastic cells. SIZE: 1193 amino acids; 130976 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. Gamma-2 is a subunit of laminin-5 (epiligrin/kalinin/nicein). SUBCELLULAR LOCATION: Secreted, extracellular space, extracellular matrix, basement membrane. Note=Major component. TISSUE SPECIFICITY: The large variant is expressed only in specific epithelial cells of embryonic and neonatal tissues. In 17-week old embryo the small variant is found in cerebral cortex, lung, and distal tubes of kidney, but not in epithelia except for distal tubuli.DOMAIN:SwissProt: Q13753 The alpha-helical domains I and II are thought to interact with other laminin chains to form a coiled coil structure. & Domain IV is globular. DISEASE: SwissProt: Q13753 # Defects in LAMC2 are a cause of junctional epidermolysis bullosa gravis (JEB) [MIM:226700]; also known as junctional epidermolysis bullosa Herlitz-Pearson type. JEB is a blistering disorder in skin that is characterized by a separation of basal cells from the basement membrane due to a decreased number of hemidesmosomes. Laminin-5 is missing from the basement membrane of patients with the gravis form of epidermolysis bullosa. SIMILARITY: Contains 8 laminin EGF-like domains. & Contains 1 laminin IV type A domain. MISCELLANEOUS: Binds heparin (By similarity). |
| Molecular Weight | 150 kDa (precursor form) and 105 kDa (mature form) |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blot on recombinant Laminin-5. Western Blot Analysis: A 1:500-1:2000 dilution of this antibody detected Laminin-5, gamma2 chain in 10 µg of human recombinant Laminin-5. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Maintain for 1 year at 2–8°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |