In undiluted form or diluted to working concentration, this material may be stored at -20°C in aliquots up to the recommended retest date for the specific lot. This data is published in the Certificate of Analysis. Avoid repeated freeze/thaw cycles. The solution may be transferred to smaller vials in working volumes for long-term storage. Plastic vials are recommended as this protein tends to adhere to glass. Please see the link below to access the product datasheet for additional details:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/409/686/l2020pis-ms.pdf
L2020
Mouse Laminin
from Engelbreth-Holm-Swarm murine sarcoma basement membrane, 0.2 μm filtered, liquid, 1-2 mg/mL, suitable for cell culture
Laminin from Engelbreth-Holm-Swarm murine sarcoma basement membrane
Synonym(e):
Laminin
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Über diesen Artikel
Biological source:
mouse (Engelbreth-Holm-Swarm mouse sarcoma basement membrane)
Form:
aqueous solution
Technique(s):
cell culture | mammalian: suitable
Concentration:
1-2 mg/mL in Tris-buffered saline
Mol wt:
A subunit 400 kDa, B1 subunit 210 kDa, B2 subunit 200 kDa
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Unterstützung erhaltenProduktname
Laminin aus Engelbreth-Holm-Swarm murine sarcoma basement membrane, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
biological source
mouse (Engelbreth-Holm-Swarm mouse sarcoma basement membrane)
product line
BioReagent
form
aqueous solution
mol wt
A subunit 400 kDa, B1 subunit 210 kDa, B2 subunit 200 kDa
packaging
pkg of 1 mg
concentration
1-2 mg/mL in Tris-buffered saline
technique(s)
cell culture | mammalian: suitable
surface coverage
1‑2 μg/cm2
impurities
Microbial Contamination, passes test
NCBI accession no.
binding specificity
Peptide Source: Collagen
shipped in
dry ice
storage temp.
−20°C
Quality Level
Gene Information
mouse ... Lama1(16772), Lamb1(16777), Lamc1(226519), Lamc2(16782)
Verwandte Kategorien
1 of 4
Dieser Artikel | |||
|---|---|---|---|
| technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable |
| biological source mouse (Engelbreth-Holm-Swarm mouse sarcoma basement membrane) | biological source human placenta | biological source mouse (Engelbreth-Holm-Swarm (EHS) sarcoma) | biological source human plasma |
| concentration 1-2 mg/mL in Tris-buffered saline | concentration 0.5 mg/mL in Tris-buffered saline | concentration - | concentration - |
| Quality Level 300 | Quality Level 300 | Quality Level 100 | Quality Level 300 |
| mol wt A subunit 400 kDa, B2 subunit 200 kDa, B1 subunit 210 kDa | mol wt - | mol wt 900 kDa | mol wt A subunit 75 kDa, B subunit 65 kDa |
| form aqueous solution | form liquid | form solution | form lyophilized powder |
General description
Laminine sind kollagenänhliche Glykoproteine[1] mit einem erheblichen Molekulargewicht. Es handelt sich dabei um heterotrimere Proteine mit drei Disulfid-gebundenen Polypeptiden, die als α-, β- und γ-Ketten bezeichnet werden.[2] Die multiplen Polypeptiddomänen von Lamininen können an verschiedene Zelloberflächenrezeptoren binden, darunter mehrere Integrin-Dimere, Dystroglycan und Syndecan sowie andere Komponenten der EZM. Dieses trimere Glykoprotein der extrazellulären Matrix bildet den Hauptbestandteil der Basallamina, welche die Adhäsion, Proliferation und Differenzierung fördert.
Application
Laminin aus der Basalmembran von Engelbreth-Holm-Swarm-Maus-Sarkomzellen wurde in folgenden Anwendungen eingesetzt:
Laminin unterstützt das Wachstum und die Differenzierung vieler Zelltypen, darunter Epithel-, Endothel-, Nerven-, Muskel- und Leberzellen. Es wird zur Verwendung als Trägerschicht (Substratum) in der Zellkultur empfohlen (1-2 μg/cm2). Die optimale Konzentration hängt vom Zelltyp und auch von der Anwendung oder den Forschungszielen ab.
Biochem/physiol Actions
Lamininproteine sind integrale Bestandteile des strukturellen Gerüsts von tierischen Geweben. Sie verbinden sich mit Kollagen Typ IV über Entactin und Perlecan und binden durch Integrinrezeptoren, Dystroglykan-Glykoprotein-Komplexe und Lutheran-Blutgruppenglykoproteine an die Zellmembranen. Laminin verfügt über aktive Domänen für die Kollagenbindung, Zelladhäsion, Heparinbindung und das Neuritenauswuchs-Fragment.
Preparation Note
Dieses Produkt wird in einer Konzentration von 1 − 2 mg/ml in 50 mM Tris-HCl mit 150 mM NaCl geliefert. Es wurde durch einen 0,2-μm-Filter filtriert. Lösung vor Gebrauch langsam bei 2–8°C auftauen, um Gelbildung zu verhindern. Für die Verwendung als Beschichtung in einer ausgeglichenen Salzlösung verdünnen, die Kulturoberfläche mit dem kleinstmöglichen Volumen beschichten und zwei Stunden bei 37 °C inkubieren. Vor dem Ausplattieren der Zellen dreimal mit PBS waschen. Lamininbeschichtungen können einen Monat bei 2–8 °C gelagert werden.
Other Notes
Laminin ist ein trimeres Glykoprotein der extrazellulären Matrix mit vielen Domänen und ist neben dem Kollagen der Hauptbestandteil der Basallamina, die die Adhäsion, Proliferation und Differenzierung fördert. Laminin besteht aus A-, B1- und B2-Ketten, die über viele Disulfidbindungen verknüpft sind. Dieses Lamininprodukt wurde aus Engelbreth-Holm-Swarm-Maustumoren isoliert.
Disclaimer
Es wird empfohlen, dieses Produkt in Arbeitsaliquoten bei -20 °C zu lagern.
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Protokolle
Die Beschichtung von Oberflächen mit Laminin für die Zellkultivierung erfordert besondere Bedingungen, damit optimale Ergebnisse erzielt werden. Enthalten sind Protokolle für die Beschichtung von Deckgläsern für die Kultur von Neurosphären und für die allgemeine Zellkultur.
Coating surfaces with laminin for culturing cells requires specific conditions for optimal results. Protocols for coating coverslips to culture neurospheres and general cell culture are included.
A stem cell culture protocol to generate 3D NSC models of Alzheimer’s disease using ReNcell human neural stem cell lines.
Step-by-step culture protocols for neural stem cell culture including NSC isolation, expansion, differentiation and characterization.
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How long can stocks be frozen at -20C for?
1 answer-
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How will the product L2020 shipped? Will it be shipped in room temperature, on ice, or on dry ice?
1 answer-
The product L2020 will be shipped using dry ice. Upon receipt, it should be stored at -20 degrees Celsius.
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How is this laminin concentration determined? (by which method?) I tried to measure the concentration of this product by ELISA, and the results always came out half of what is written on the vial.. what is the protein purity?
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Laminin concentration is 1.0 - 2.0 mg protein/ml. The protein content is determined by Lowry method. Please refer the attached document for more details. https://www.sigmaaldrich.com/specification-sheets/131/942/L2020-1MG_________SIGMA____.pdf
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Can I coat Laminin, Product L2020, on top of poly-L-lysine for better adherence?
1 answer-
When coating coverslips or slides for use with neural stem cells (neurospheres), the following procedure is recommended. Precoat the coverslips with poly-L-lysine (Product No. P5899) at 100 μg/mL. This is incubated for 30 minutes or more at room temperature. The coverslips are then washed with sterile water and allowed to dry. The poly-lysine coated coverslips can be stored at room temperature until ready to use (up to ~1 month). Coverslips are then coated with 30 - 50 μL of warmed laminin, which is at a concentration of 50 μg/mL. Incubate at 37°C for 30 minutes. Remove the laminin, and wash with DME (50 μL) twice. Cells are then added to the coverslips.
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How can I store solutions of Laminin, Product L2020?
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The product has a recommended storage temperature of -20°C. Please note, the product needs to be thawed slowly at 2-8°C. If the product has been warmed up at room temperature and it gels, it cannot be reactivated for use. We recommend storage at 2-8°C for up to one week. For longer storage, the product needs to be frozen at -20°C in single-use aliquots.
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What is the protein concentration of the solution of Product L2020, Laminin?
1 answer-
The protein specification for L2020 is 1.0 to 2.0 mg/mL. The exact value can be found on the lot-specific Certificate of Analysis, which can be downloaded from our website.
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What is the purity of Laminin, Product L2020?
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We do not determine a purity of this product. An SDS PAGE gel is run and the pattern (3 bands) is comparable to past lots.
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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What type of laminin is Product L2020?
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In the nomenclature system by Burgeson, this is Laminin 1. The laminins were numbered with Arabic numerals in the order discovered. The previous A, B1, and B2 chains, and their isoforms are listed by the Greek letters, for alpha, beta, and gamma, respectively, followed by an Arabic numeral to identify the isoforms. For example, the first laminin identified from the Engelbreth-Holm-Swarm tumor (EHS) was designated laminin-1 with the chain composition α-1/β-1/γ-1. The genes for these 3 chains are LAMA1, LAMB1, and LAMC1. Reference: Burgeson, R.E., et al., A new nomenclature for the laminins. Matrix Biol., 14, 209-211 (1994).
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