anti mac 1 rabbit antibody  (Bio-Rad)

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Activated Protein C Accelerates Venous Thrombus Resolution through Heme Oxygenase-1 Induction

doi: 10.1111/jth.12424

Figure Lengend Snippet: Venous thrombi were taken from APC- or vehicle-treated mice on Day 7 and 12 post-surgery. A) IL-6 concentrations within the thrombus homogenates were measured by ELISA and normalized to the total amount of protein in the lysates. Data shown are the means ± SEM. *, p<0.05, vehicle vs. APC, n=5-8. B) Collagen deposition was analyzed by staining 5μm-thick paraffin sections of the venous thrombi with Masson’s Trichrome Stain. Quantification was done by NIH ImageJ of 2-4 mice per group. C) Macrophage accumulation was analyzed by immunohistochemical staining with an anti-Mac-2 antibody. Scale bars: 50 μm.

Article Snippet: Two-color immunohistochemical staining of the paraffin sections was done using the Multiple Antigen Labeling kit from Vector Laboratories with a rabbit anti-HO-1 antibody and a rat anti-Mac-2 antibody (for macrophages), based on our published methods [ 13 ].

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemical staining

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Activated Protein C Accelerates Venous Thrombus Resolution through Heme Oxygenase-1 Induction

doi: 10.1111/jth.12424

Figure Lengend Snippet: A) Mice were injected daily with vehicle or APC from Day 4 to Day 11 following IVC ligation. Intra-thrombus HO-1 levels on Day 7 and Day 12 were determined by immunoblot analysis using an anti-HO-1 antibody. The amount of HO-1 was quantified by NIH ImageJ. Data shown are the means ± SEM. *, p= 0.0181, vehicle vs. APC, n=3-5. B) Bone marrow-derived macrophages were treated with LPS and IFN-γ, with or without APC (5μg/ml) for 12 hours. HO-1 gene transcription was quantified by real-time qRT-PCR. Changes in HO-1 expression were calculated using the 2−ΔCt method (RQ Manager 1.4). The expression of 36B4 gene in the same cDNA set was used as a control. Data shown are the means ± SD of two independent experiments. C) The 5μm-thick paraffin sections of Day 12 venous thrombi from control and APC-treated mice were subjected to immunohistochemical staining with anti-HO-1 (in red) and anti-Mac-2 (macrophages; in brown/gray). Arrows indicate HO-1/Mac-2 double-positive cells and arrowheads indicate HO-1 or Mac-2 single-positive cells.

Article Snippet: Two-color immunohistochemical staining of the paraffin sections was done using the Multiple Antigen Labeling kit from Vector Laboratories with a rabbit anti-HO-1 antibody and a rat anti-Mac-2 antibody (for macrophages), based on our published methods [ 13 ].

Techniques: Injection, Ligation, Western Blot, Derivative Assay, Quantitative RT-PCR, Expressing, Immunohistochemical staining, Staining