Immunohistochemical staining was performed on 4-μm formalin-fixed, paraffin-embedded skin sections. Endogenous peroxidase activity was blocked with 3% hydrogen peroxide (Dako Cytomation GmbH, Hamburg, Germany). Antigen retrieval was carried out in Tris-EDTA buffer (1mM, pH 8.5) for 5 min at 120°C in a pressure cooker. Ki67 [SP6] antibody (abcam, Cambridge, UK) at a dilution of 1:500 and murine ß-defensin-2 antibody (DEFB4 ABIN674574 from antibodies-online) at a dilution of 1:100 was applied for 30 min at room temperature. Subsequent reactions were performed with the Histofine Simple Stain Mouse MAX PO (rabbit) biotin-free horseradish peroxidase enzyme–labeled polymer detection system. Positive reactions were visualized with diaminobenzidine (DAB) (Dako) followed by counterstaining with hematoxylin (Merk, Darmstadt, Germany). Evaluation of the staining was performed semi-quantitatively by means of light microscopy (Carl Zeiss Vision, Hallbergmoos, Germany) in magnification as indicated.
Ki67 sp6 antibody
Manufactured by Abcam
Sourced in United Kingdom
The Ki67 [SP6] antibody is a laboratory reagent used to detect the presence and expression of the Ki67 protein, which is a cellular marker for proliferation. This antibody is a recombinant monoclonal antibody that specifically binds to the Ki67 protein in various cell types. The Ki67 protein is expressed during all active phases of the cell cycle (G1, S, G2, and mitosis), but is absent in resting cells (G0). The Ki67 [SP6] antibody can be utilized in various laboratory techniques, such as immunohistochemistry and flow cytometry, to study cell proliferation in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Hydrogen peroxide, by Agilent Technologies (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
Diaminobenzidine dab, by Agilent Technologies (1 mentions)
3 3 diaminobenzidine tetrahydrochloride, by Dojindo Laboratories (1 mentions)
Vectastain elite abc, by Vector Laboratories (1 mentions)
Apoptag peroxidase in situ apoptosis detection kit, by Merck Group (1 mentions)
Itraq reagent 4 plex kit, by AB Sciex (1 mentions)
3 protocols using ki67 sp6 antibody
1
Immunohistochemical Staining of Skin Sections
2
Apoptosis Detection and Protein Quantification
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3, 3'-diaminobenzidine tetrahydrochloride was obtained from Dojindo and dextran sodium sulfate (DSS) from ICN Biomedicals (Aurora, OH). An ApopTag® Peroxidase In Situ Apoptosis Detection Kit was purchased from Millipore and an iTRAQ Reagent 4 Plex Kit from AB Sciex. The rabbit monoclonal Ki-67 [SP6] antibody was the product of Abcam. VECTASTAIN Elite ABC was purchased from Vector Laboratories Ltd.
3
Fibroblast Proliferation Analysis
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Proliferation analysis on 1 Â 10 6 formalin-fixed and embedded fibroblasts (passage 2) of wt and BMP6 e/e mice was done with the Shandon Cytoblock Cell Block Preparation System (Thermo Electron Corporation, Pittsburgh, PA) and the subsequent Ki67 IH analysis with Ki67 (SP6) antibody (1:500; Abcam, Cambridge, UK). Ki67-positive fibroblasts were counted from six fields of views, averaged, and were normalized to total cell counts.
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