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Anti laminin α5

Manufactured by Abcam

Anti-Laminin α5 is a primary antibody that recognizes the alpha 5 subunit of the laminin protein. Laminin is a major component of the extracellular matrix and plays a crucial role in cell adhesion, migration, and differentiation. This antibody can be used in various research applications that involve the study of laminin and its interactions with cells.

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2 protocols using anti laminin α5

1

Western Blot Analysis of Palatal Proteins

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RIPA buffer was used to extract total protein from the palatal explants. A BCA Protein Assay Kit (Solarbio, China) was then used to detect protein content in the supernatant. The protein was denatured at 100 °C for 5 min, then (∼20 μg) electrophoresed on SDS-PAGE gel and transferred onto a PVDF membrane (Merck Millipore, USA). The membranes were blocked with 5% skimmed milk in Tris-buffered saline containing 0.05% Tween-20 and incubated with primary antibodies at 4 °C overnight. The antibodies used were anti-Laminin α5 (Abcam, AB184330, 1:1000), anti-ki67 (Affinity, AF0931, 1:1000), anti-cyclin D1 (Affinity, AF0931, 1:1000), anti-cleaved caspase 3 (Cell Signaling Technology, 9664, 1:1000), anti-gli1 (Affinity, AF0931, 1:1000), anti-β-actin (Elabscience, E-AB-20058, 1:1000). Then the membranes were incubated with a secondary antibody at room temperature for 1.5 h. An ECL kit (Cell Signaling Technology, USA) was used to detect the protein bands following the manufacturer's protocols. β-actin was used as the load control. The grey level was quantified using ImageJ v1.8.0 software (National Institutes of Health).
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2

Immunostaining of Muscle Sections

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For immunostaining, 10μm cryostat cut frozen muscle sections were blocked in 5% goat serum in PBS at room temperature for 1 hour. Sections were then incubated with anti-Laminin (Sigma Aldrich; St. Louis, MO) at 1μg/ml, at room temperature for 2 hours, or with one of the following antibodies: anti-Utrophin (Novocastra, Buffalo Grove, IL) at 7.5 μg/ml, anti-Plectin 1 (Santa Cruz, Dallas, TX) at 1 μg/ml, anti-Laminin α5 (Abcam, Cambridge, MA) at 10 μg/ml, anti-α-dystroglycan (clone IIH6C4, Sigma Aldrich, St. Louis, MO) at 1:100, or anti-dystroglycan (R&D Systems, Minneapolis, MN) at 5 μg/ml at 4° C overnight. Sections were incubated in appropriate Cy3-conjugated secondary antibodies (Jackson Immunoresearch; West Grove, PA) at 7.5μg/mL and FITC-conjugated Wisteria floribunda agglutinin (WFA, Vector Laboratories, Burlingame, CA) at 1μg/ml for 1 hour at room temperature and then mounted using ProLong Gold Antifade Mountant (Invitrogen, Carlsbad, CA).
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