Primary rat alveolar macrophages were isolated and plated on a 16-well chamber slide as previously described24 (link) prior to treatment with or without 5 nM of MMP-9 inhibitor. Wells were then fixed with 4% paraformaldehyde and incubated with polyclonal rabbit antibodies to MMP-9 (Abcam) or receptor for advanced glycation end (RAGE) antibody (Abcam, Waltham, MA/USA) overnight at 4°C prior to incubation with an anti-rabbit Alexa Fluor 488 secondary antibody (Jackson ImmunoResearch, West Grove, PA/USA) and DAPI (Vector Laboratories, Burlingame, CA/USA) for nuclear staining. Images of 80–100 alveolar macrophages / experimental group were acquired with an Olympus fluorescent microscope (Center Valley, PA/USA). Random fields were selected from each slide and at least five images were acquired per sample for a total of 80–100 cells/condition. ImageJ (NIH, Bethesda, MD/USA) was used to perform automated background correction and to process the images into a binary image map to allow for automated cell border acquisition and assessment of mean fluorescence intensity.
Polyclonal rabbit antibodies to mmp 9
Manufactured by Abcam
Sourced in United States
Polyclonal rabbit antibodies to MMP-9 are laboratory reagents produced in rabbits that recognize the matrix metalloproteinase 9 (MMP-9) protein. MMP-9 is an enzyme involved in the breakdown of extracellular matrix components.
Automatically generated - may contain errors
2 protocols using polyclonal rabbit antibodies to mmp 9
1
MMP-9 Inhibition and RAGE Expression in Rat Alveolar Macrophages
2
Quantification of MMP-9 and RAGE in Rat Alveolar Macrophages
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Primary rat alveolar macrophages were isolated and plated on a 16-well chamber slide as previously described24 (link) before treatment with or without 5 nM of MMP-9 inhibitor. Wells were then fixed with 4% paraformaldehyde and incubated with polyclonal rabbit antibodies to MMP-9 (Abcam) or receptor for advanced glycation end (RAGE) antibody (Abcam, Waltham, MA) overnight at 4 °C before incubation with an antirabbit Alexa Fluor 488 secondary antibody (Jackson ImmunoResearch, West Grove, PA) and DAPI (Vector Laboratories, Burlingame, CA) for nuclear staining. Images of 80–100 alveolar macrophages/experimental group were acquired with an Olympus fluorescent microscope (Center Valley, PA). Random fields were selected from each slide and at least 5 images were acquired per sample for a total of 80–100 cells/condition. ImageJ (NIH, Bethesda, MD) was used to perform automated background correction and to process the images into a binary image map to allow for automated cell border acquisition and assessment of mean fluorescence intensity.
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