Lamc1

Manufactured by Merck Group

LAMC1 is a specialized laboratory equipment designed for various research and analytical applications. It serves as a versatile tool for conducting a range of experiments and analyses. The core function of LAMC1 is to provide accurate and reliable measurements and data collection, supporting scientific investigations across different fields.

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Lab products found in correlation

Any kd gel, by Bio-Rad (2 mentions) Chemidoc mp, by Bio-Rad (2 mentions) Dako antibody diluent, by Agilent Technologies (1 mentions) Human ki67, by Agilent Technologies (1 mentions) Phospho histone h3 ser10 antibody, by Cell Signaling Technology (1 mentions) Liquid dab, by Agilent Technologies (1 mentions)

3 protocols using lamc1

Epitope-Tagged SFTPA2 Expression Construct

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The expression constructs for epitope‐tagged SFTPA2 were described previously (Wang et al., 2009 (link)). A gBlock with each of the variants we identified was synthesized at IDT and then incorporated into the expression plasmid using a Gibson Assembly (Gibson et al., 2009 (link)). Each plasmid was sequence verified. A549 cells were acquired from the American Type Culture Collection (ATCC) and propagated in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and penicillin, streptomycin, and l‐glutamine. Cells were transfected with Fugene HD Transfection reagent (Roche). Forty‐eight hours after transfection, cells and supernatant were collected for Western blot analysis. Fifteen microliters of supernatant was mixed directly with loading buffer and 20 μg of cell lysates, extracted in radioimmunoprecipitation assay buffer, were separated on Any kD gels (Bio‐Rad) and blotted using standard procedures. Antibodies for V5 (Thermo Fisher Scientific), LAMC1 (Sigma‐Aldrich), and Tubulin (Bio‐Rad) were used at 1:1000 overnight before imaging (Chemidoc MP; Bio‐Rad).

Sutton R.M., Bittar H.T., Sullivan D.I., Silva A.G., Bahudhanapati H., Parikh A.H., Zhang Y., Gibson K., McDyer J.F., Kass D.J, & Alder J.K. (2022). Rare surfactant‐related variants in familial and sporadic pulmonary fibrosis. Human Mutation, 43(12), 2091-2101.

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Epitope-tagged SFTPA2 Variant Expression

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The expression constructs for epitope-tagged SFTPA2 were described previously (Wang et al., 2009 (link)). A gBlock with each of the variants we identified was synthesized at IDT and then incorporated into the expression plasmid using a Gibson Assembly (Gibson et al., 2009 (link)). Each plasmid was sequence verified. A549 cells were acquired from the American Type Culture Collection (ATCC) and propagated in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and penicillin, streptomycin, and l-glutamine. Cells were transfected with Fugene HD Transfection reagent (Roche). Forty-eight hours after transfection, cells and supernatant were collected for Western blot analysis. Fifteen microliters of supernatant was mixed directly with loading buffer and 20 μg of cell lysates, extracted in radioimmunoprecipitation assay buffer, were separated on Any kD gels (Bio-Rad) and blotted using standard procedures. Antibodies for V5 (Thermo Fisher Scientific), LAMC1 (Sigma-Aldrich), and Tubulin (Bio-Rad) were used at 1:1000 overnight before imaging (Chemidoc MP; Bio-Rad).

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Immunohistochemical Staining of Paraffin Sections

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Paraffin-embedded tissue sections (4 μm) were deparaffinized in xylene and rehydrated in graded alcohols. Antigen retrieval was performed by incubating tissue sections with TrilogyTM (Cell Marque, Austin, TX, catalog # CMX833). Endogenous peroxidase activity was blocked by incubation with 3% H₂O₂ for 15 min. Sections were pre-incubated with Dako Antibody Diluent (Dako, Carpinteria, CA) at room temperature for 30 min, followed by incubation with antibody diluted in Dako Antibody Diluent at 4°C overnight. Positive reactions were detected by applying EnVision™+/HRP polymer (Dako) for 30 min, followed by incubation in DAB substrate for 5 min (Liquid DAB+, Dako). The slides were then counterstained with hematoxylin to visualize the cell nuclei. Antibodies used were: human Ki-67 (cat # M7240, Dako), mouse Ki-67 (cat # 12202, Cell Signaling Technology), LAMC1 (cat # HPA001908, Sigma-Aldrich), and phospho-Histone H3 (Ser10) antibody (cat # 9701, Cell Signaling Technology).

Kobayashi Y., Kashima H., Wu R.C., Jung J.G., Kuan J.C., Gu J., Xuan J., Sokoll L., Visvanathan K., Shih I.M, & Wang T.L. (2015). Mevalonate Pathway Antagonist Inhibits Proliferation of Serous Tubal Intraepithelial Carcinoma and Ovarian Carcinoma in Mouse Models. Clinical cancer research : an official journal of the American Association for Cancer Research, 21(20), 4652-4662.

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