Adult mice (6–10 weeks of age) were anaesthetized and shaved, and four full-thickness wounds were made on the back of each mouse using a sterile 4-mm biopsy punch, as described (Mitchell et al., 2009 (link)). After allowing wounds to heal for 10 days, mice were euthanized by CO2 narcosis and wounds were surgically excised. Wounds were frozen in OCT compound, and 10 µm sections were prepared for immunohistology (Albany Medical College Histology Services). For immunostaining, frozen sections were rehydrated in PBS with 0.2% Tween-20 for 10 minutes, blocked in 10% heat-inactivated goat serum and 5% milk in PBS for 1 hour, then stained with the following rabbit polyclonal antisera: anti-LN-332 (1:200; Abcam, Cambridge, MA); anti-LNγ2 L4m (1:1000) (Sasaki et al., 2001 (link)); or anti-BMP-1 (1:100; Abcam, Cambridge, MA). The anti-BMP-1 antibody used here was derived from a synthetic peptide based on the carboxyterminal end of the mTLD form of BMP-1, and is likely specific to mTLD. The secondary antibody used was Alexa Fluor 594 goat anti-rabbit IgG (1:250; Molecular Probes, Eugene, OR). Images were collected on a Nikon Eclipse 80i using a Spot camera (Diagnostic Instruments, Sterling Heights, MI).
Anti ln 332
Manufactured by Abcam
Anti-LN-332 is a primary antibody that recognizes the laminin-332 protein, also known as laminin-5. Laminin-332 is a protein found in the extracellular matrix and plays a role in cell adhesion and migration. This antibody is suitable for use in various laboratory applications, such as immunohistochemistry and Western blotting, to detect and localize the laminin-332 protein.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 594 goat anti rabbit igg, by Thermo Fisher Scientific (4 mentions)
Anti lnγ2l4m, by Abcam (3 mentions)
Anti cytokeratin 14, by Abcam (2 mentions)
Anti entactin nidogen, by Abcam (2 mentions)
Anti bmp 1, by Abcam (1 mentions)
Alexa fluor 594 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Anti fn, by Merck Group (1 mentions)
Alexa fluor 488 goat anti rat igg, by Thermo Fisher Scientific (1 mentions)
Anti mmp 9, by Merck Group (1 mentions)
Anti keratin 14, by Fortrea (1 mentions)
Anti cd31, by BD (1 mentions)
4 protocols using anti ln 332
1
Immunohistological Analysis of Wound Healing
2
Immunofluorescence Staining of Basement Membrane Proteins
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Frozen sections (10μM) were rehydrated in PBS with 0.2% Tween-20 for 10 minutes, blocked in 10% heat-inactivated goat serum and 5% milk in PBS for 1 hour, then stained with the following rabbit polyclonal antisera: anti-α3 integrin or corresponding pre-immune serum (1:100)(DiPersio et al., 1995 (link)); anti-LN-332 (1:200; Abcam, Cambridge, MA); anti-entactin/nidogen (1:1000; Abcam); anti-LNγ2L4m (1:1000)(Sasaki et al., 2001 (link)); anti-fibulin-2 (1:2000)(Pan et al., 1993 (link)). Sections were co-immunostained in some cases with mouse monoclonal anti-cytokeratin 14 (1:500; Abcam). Secondary antibodies were fluorescein-conjugated goat anti-mouse IgG (1:250; Pierce, Rockford, IL) or Alexa Fluor 594 goat anti-rabbit IgG (1:250; Molecular Probes, Eugene, OR), as appropriate. Images were collected on a Nikon Eclipse 80i using a Spot camera (Diagnostic Instruments, Sterling Heights, MI).
3
Immunofluorescence Staining of Wound Tissue
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10-µm frozen tissue sections were rehydrated in 0.2% Tween 20/PBS for 10 min, fixed in 3.7% formaldehyde, permeabilized in 0.5% Triton X-100, blocked in 10% heat-inactivated goat serum/5% milk/PBS for 1 h, and then stained with anti–cleaved caspase 3, anti-CD31 (BD), anti–LN-332 (Abcam), anti-FN (Sigma-Aldrich), anti–MMP-9 (Sigma-Aldrich), or anti–keratin 14 (Covance). Immunostaining was also performed using antibodies against integrin α2 (CD49b; EMD Millipore), α5 (5H10-27; BD), and α6 (GoH3; EMD Millipore). Secondary antibodies were Alexa Fluor 488 goat anti–rat IgG, Alexa Fluor 488 goat anti–hamster IgG, Alexa Fluor 594 goat anti–mouse IgG, or Alexa Fluor 594 goat anti–rabbit IgG (Molecular Probes), as appropriate. Images were collected on an Eclipse 80i upright microscope using a Spot camera. For assessment of wound immunostaining, the field within the wound bed below the reepithelialized epidermis was imaged. For assessment of vessel density within papillomas, nonnecrotic tumor regions were imaged. CD31 and MMP-9 staining was quantified using ImageJ software.
4
Immunofluorescence Staining of Basement Membrane Proteins
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