The following primary antibodies were used on paraffin-embedded sections: anti α-SMA (Clone 1A4, Code M0851, dilution 1:100, Dakocytomation, Glostrup, Denmark), anti-CD62P (P-selectin, NCL-CD62P-367, dilution 1:50, Novocastra Laboratories Ltd, Newcastle upon Tyne, United Kingdom), anti-CD68 (Clone PG-M1, code M0876, dilution 1:100, Dakocytomation, Glostrup Denmark), anti-CD8 (Clone CD8/144B, code M7103, dilution 1:50, Dakocytomation, Glostrup, Denmark) and anti-CD4 (NCL-L-CD4–1F6, dilution 1:100, Novocastra Laboratories Ltd, Newcastle upon Tyne, United Kingdom). To avoid unspecific binding slides were pre-incubated with the Rodent Block of the Mouse on Mouse Polymer Kit (Abcam, Cambridge, United Kingdom). The Vectastain Elite ABC Kit Universal (Vector Laboratories, Burlingame, Ca, USA) was applied according to the instructions. The activity of endogenous peroxidase was blocked with 3% hydrogen peroxide (H2O2) in methanol. The sections were visualised with the Vector DAB Substrate Kit for Peroxidase (SK-4100; Vector Laboratories Inc., Burlingame, California, USA) and counterstained with haematoxylin. P-selectin as well as α-SMA were assessed based on a previously established four-graded classification63 (link)64 (link). A blinded scientist performed semiquantitative analysis of cell infiltration in 5 non-overlapping high power fields (HPF).
Ncl l cd4 1f6
Manufactured by Leica Biosystems
Sourced in Denmark, Germany
The NCL-L-CD4–1F6 is a monoclonal antibody that can be used to detect the CD4 antigen in immunohistochemical staining procedures. It is intended for laboratory use.
Automatically generated - may contain errors
Lab products found in correlation
Vectastain universal elite abc kit, by Vector Laboratories (1 mentions)
Autostainer, by Leica Biosystems (1 mentions)
M7103, by Agilent Technologies (1 mentions)
M7158, by Agilent Technologies (1 mentions)
M0888, by Agilent Technologies (1 mentions)
M7050, by Agilent Technologies (1 mentions)
M7240, by Agilent Technologies (1 mentions)
2 protocols using ncl l cd4 1f6
1
Immunohistochemical Analysis of Tissue Markers
2
Autopsy Examination Protocol
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An autopsy was performed two hours after the patient’s death. No craniotomy was performed. Written informed consent was obtained from the patient’s family before autopsy. The protocol for reporting this case was approved by the ethics committee of Asahikawa Medical University (approval number #17115, August 28, 2017). The systemic organs were removed, examined, and fixed in phosphate-buffered 10 % formalin. Tissues were processed to paraffin blocks and the Section (4 μm) were subjected to hematoxylin and eosin staining, as well as Berlin blue staining. An autostainer (Leica Biosystems, Nußloch, Germany) was used to perform immunohistochemistry assessments of CD79a (antibody: M7050, DAKO, Carpinteria, CA), CD4 (antibody: NCL-L -CD4-1F6, Novocastra, Leica Microsystems, Wetzlar, Germany), CD8 (antibody: M7103, DAKO), HepPar1 (antibody: M7158, DAKO), cytokeratin 19 (CK19) (antibody: M0888, DAKO), and Ki-67 (MIB-1) (M7240, DAKO) expression.
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