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Laminin 1 2

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Laminin 1-2 is a component of the extracellular matrix (ECM) that plays a crucial role in cell adhesion, migration, and differentiation. It is a heterotrimer composed of α1, β1, and γ1 chains.

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Lab products found in correlation

Tissue plus o c t compound, by Thermo Fisher Scientific (3 mentions) Lsm 710 confocal microscope, by Zeiss (3 mentions) Enhanced chemiluminescence plus system, by Cytiva (1 mentions) Fluorchem hd2 camera, by Cell Biosciences (1 mentions) Fluorchem software, by Cell Biosciences (1 mentions) Polyvinylidene difluoride membrane, by Cytiva (1 mentions) Coomassie blue, by Bio-Rad (1 mentions) Bradford assay, by Bio-Rad (1 mentions) Prolong diamond antifade mountant, by Thermo Fisher Scientific (1 mentions) Pdgfrα, by R&D Systems (1 mentions) P smad3, by Novus Biologicals (1 mentions) Tgf β1, by Novus Biologicals (1 mentions) Perilipin 1, by Abcam (1 mentions) F4 80, by Abcam (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Superfrost plus slide, by Thermo Fisher Scientific (1 mentions)

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Laminin

3 protocols using laminin 1 2

1

Western Blot Analysis of PFC Proteins

Cited 1 time
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Western immunoblotting was conducted as previously described [14 (link), 33 (link), 76 (link)]. In brief, around 50 mg of PFC tissues were sonicated in ice-cold 1% SDS and immediately boiled. Protein concentrations were ascertained using the Bradford assay (BioRad, Hercules, CA). Equal amounts of protein (10-30 μg) were separated by SDS-PAGE into slab gels of 10-15% polyacrylamide and transferred to polyvinylidene difluoride membranes (Amersham Biosciences, Baie d'Urfé, Quebec). The membranes were incubated with primary antibodies against APE1, OGG1, DNMT1, DNMT3A, MeCP2, BMP4, DNMT3B, Laminin 1-2 (1:1000, Abcam), and actin (1:2000, Abcam) overnight at 4°C. Primary antibody binding was detected using horseradish peroxidase-conjugated secondary antibodies and the Enhanced Chemiluminescence Plus System (Amersham Biosciences, Baie d'Urfé, Quebec). Chemiluminescence was detected using a FluorChem HD2 camera with FluorChem software (Cell Biosciences). The membranes were stained with Coomassie blue (BioRad, Hercules, CA) to confirm equal protein loading. Signals were quantified using NIH Image J64 software and normalised relative to actin or Coomassie staining.
Kovalchuk A., Ilnytskyy Y., Rodriguez-Juarez R., Shpyleva S., Melnyk S., Pogribny I., Katz A., Sidransky D., Kovalchuk O, & Kolb B. (2017). Chemo brain or tumor brain - that is the question: the presence of extracranial tumors profoundly affects molecular processes in the prefrontal cortex of TumorGraft mice. Aging (Albany NY), 9(7), 1660-1675.
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2

Immunofluorescent Analysis of Splenic B Cells

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Spleens were collected from recipient mice at different time points after B cell transfer, and 5 mm fragments were fixed for 4 hours in PBS with 4% paraformaldehyde and then overnight in PBS with 30% sucrose. Tissues were cryopreserved in Tissue-Plus OCT compound (Thermo Fisher Scientific), and 10 μm sections were obtained using a cryostat. Tissue sections were hydrated and then blocked for 30 minutes at room temperature in staining solution (5% donkey serum in PBS-Tween 20) plus FcR block (clone 93; BioLegend). Antibody staining was performed overnight at 4°C in staining solution with 1–2 ng/μL of the following anti-mouse antibodies: anti-CD45.1-AF488 (clone A20; BioLegend), CD169-AF647 (clone 3D6.112; BioLegend), and laminin 1+2 (polyclonal rabbit anti-mouse, ab7463; Abcam) labeled with CF568-Mix-n-Stain antibody labeling kit (Biotium). Slides were washed with PBS, mounted using ProLong Diamond Antifade Mountant (Invitrogen), and imaged using 20× magnification lenses in a Zeiss LSM710 confocal microscope. Images were processed and analyzed using Volocity 6.3. software (PerkinElmer).
Gómez Atria D., Gaudette B.T., Londregan J., Kelly S., Perkey E., Allman A., Srivastava B., Koch U., Radtke F., Ludewig B., Siebel C.W., Ryan R.J., Robertson T.F., Burkhardt J.K., Pear W.S., Allman D, & Maillard I. (2022). Stromal Notch ligands foster lymphopenia-driven functional plasticity and homeostatic proliferation of naive B cells. The Journal of Clinical Investigation, 132(13), e158885.
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3

Muscle Regeneration Histologic Evaluation

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Histologic evaluation was performed at indicated time points in tibialis anterior muscles from wild type C57BL/6J (both injured and contralateral uninjured TAs), LysM-Cre;Tgfb1fl/fl mutants on a C57BL/6J background, and TGFBRII-FC treated (WT) mice following inductive ischemia/reperfusion injury and concomitant cardiotoxin at indicated timepoints. Muscle specimens were flash frozen and sectioned as described above, subsequently mounted on Superfrost plus slides (Fisher) and stored at −20°C. Sections were blocked and permeabilized with 2% serum, 1% BSA, 0.1% fish skin gelatin, 0.001% Triton X-100 and 0.0005% Tween-20. Sections were incubated overnight with the following antibodies F4/80 (Abcam), PDGFRα (R&D), TGFβ1 (Novus), pSMAD3 (Novus), Ly6G (Abcam), laminin 1/2 (Abcam), eMHC (DHSB), perilipin-1 (CST), Anti-15 Lipoxygenase 1 (ALOX15) (Abcam), and KI67 (Abcam); primary antibodies listed in table below. Fluorophore conjugated secondary antibodies were diluted to 1:200. Nuclear counterstain was performed with Hoechst 33342 (Life Technologies). Appropriate primary antibody negative controls were run simultaneously with each tested sample.
Stepien D.M., Hwang C., Marini S., Pagani C.A., Sorkin M., Visser N.D., Huber A.K., Edwards N.J., Loder S.J., Vasquez K., Aguilar C., Kumar R., Mascharak S., Longaker M.T., Li J, & Levi B. (2020). Tuning macrophage phenotype to mitigate skeletal muscle fibrosis: Macrophage and FAP Crosstalk in Muscular Polytrauma. Journal of immunology (Baltimore, Md. : 1950), 204(8), 2203-2215.
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