Ncl l cd163

Manufactured by Leica camera

Sourced in Germany

The NCL-L-CD163 is a laboratory equipment product manufactured by Leica. It is designed for use in various research and diagnostic applications. The product's core function is to facilitate the detection and analysis of the CD163 protein, which is a marker for certain cell types. This information is presented in a factual and unbiased manner, without any interpretation or extrapolation on the intended use of the product.

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Lab products found in correlation

Vectastain abc hrp kit, by Vector Laboratories (2 mentions) M7103, by Agilent Technologies (1 mentions) M0876, by Agilent Technologies (1 mentions) Vectra slide scanner, by PerkinElmer (1 mentions) Positively charged slides, by Thermo Fisher Scientific (1 mentions) Ab97959, by Abcam (1 mentions) Bond 3, by Leica Biosystems (1 mentions) Bond polymer refine detection kit, by Leica Biosystems (1 mentions) Bond rx platform, by Leica Biosystems (1 mentions)

5 protocols using ncl l cd163

Multiplex Immunofluorescence Analysis of Immune Markers

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Multiplex staining was performed per the Opal protocol staining method ^{15 (link)} for markers: CD4 (1:25, CM153BK, Biocare) with subsequent visualization using fluorescein AF-488 (1:50); CD8 (1:200, M7103, Dako) with visualization using AF-594 (1:50); CD68 (1:100, M0876, Dako) with visualization using AF-647; VISTA (1:100, Janssen) with visualization using coumarin (1:50); CD163 (1:100, NCL-L-CD163, Leica) with visualization using AF-488 and PD-L1 (1:100, 13684, Cell Signaling Technology) with visualization using AF-555 (1:50). Nuclei were visualized with DAPI (1:2000). All sections were cover-slipped using Vectashield Hardset 895 mounting media.
For multispectral analysis, each individually stained section (CD4/AF-488, CD8/AF-594, CD68/AX-647, VISTA/coumarin, PD-L1/AF-555, CD163/AF-488, and DAPI) was utilized to establish the spectral library of fluorophores. Slides were scanned using the Vectra slide scanner (PerkinElmer). For each marker, the mean fluorescent intensity per case was determined as a base point from which positive calls could be established. The co-localization algorithm was used to determine percent of PD-L1 and VISTA staining on each cellular subset. Five random areas on each sample were analyzed blindly by a pathologist at 20× magnification.

Gao J., Ward J.F., Pettaway C.A., Shi L.Z., Subudhi S.K., Vence L.M., Zhao H., Chen J., Chen H., Efstathiou E., Troncoso P., Allison J.P., Logothetis C.J., Wistuba I.I., Sepulveda M.A., Sun J., Wargo J., Blando J, & Sharma P. (2017). VISTA is an inhibitory immune checkpoint that is increased after ipilimumab therapy in patients with prostate cancer. Nature medicine, 23(5), 551-555.

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Immunohistochemistry for BTK, SOX2, and CD163

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Immunohistochemistry (IHC) staining was performed in the Pathology Department at the University of Edinburgh. Samples were sectioned from FFPE tissue blocks at a thickness of 5 μm and stained on positively charged slides (Thermo Fisher Scientific) to maximize tissue adherence. IHC was performed using a BOND III autostainer with Bond Polymer Refine Detection Kit (DS9800; Leica Biosystems) according to manufacturer instructions. Slides were blocked with Peroxide 3–4% (vol/vol) hydrogen peroxide for 5 min. Antigen retrieval was performed using Bond Epitope Retrieval Solution 1 (Citrate; pH 6.0) (AR9961; Leica Biosystems) for 20 min at 100°C. Slides were incubated at room temperature with a BTK antibody (MBS9215577, 1:100; MyBioSource), SOX2 (ab97959, 1:200; Abcam), or CD163 (NCL-L-CD163, 1:100; Leica) for 20 min. Visualisation was achieved using 3,3′-diaminobenzidine (DAB) staining and haematoxylin counterstaining (Leica Biosystems). Finally, slides were dehydrated and cleared in xylene before coverslips were applied. Three FFPE tissue blocks (GBM, non-tumour brain, and tonsil, which additionally served as a positive control) were used to optimize the BTK antibody dilutions before applying on the TMA.

Al Shboul S., Curran O.E., Alfaro J.A., Lickiss F., Nita E., Kowalski J., Naji F., Nenutil R., Ball K.L., Krejcir R., Vojtesek B., Hupp T.R, & Brennan P.M. (2021). Kinomics platform using GBM tissue identifies BTK as being associated with higher patient survival. Life Science Alliance, 4(12), e202101054.

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Immunohistochemical Profiling of NPC Samples

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The NPC tissue samples were dewaxed in xylene and ethanol and steamed in a steamer autoclave with pH 8.0 antigen retrieval solution to retrieve antigen epitopes. Endogenous peroxidase was eliminated by 3% hydrogen peroxide, followed by blocking with 10% goat serum for 1 h. The slides were incubated at 4°C overnight with a primary antibody: an anti-human PD-L1 rabbit monoclonal antibody (dilution 1:200; clone E1L3N; Cell Signaling Technology, Danvers, MA, USA), an anti-human CD68 mouse monoclonal antibody (dilution 1:200; KP-1; Agilent, Santa Clara, CA, USA), and an anti-human CD163 mouse monoclonal antibody (dilution 1:200; NCL-L-CD163; Leica, Nussloch, Germany). The secondary antibody was a ready-to-use VECTASTAIN ABC HRP Kit (Vector Laboratories, Burlingame, CA, USA).

Deng R., Lu J., Liu X., Peng X.H., Wang J, & Li X.P. (2020). PD-L1 Expression is Highly Associated with Tumor-Associated Macrophage Infiltration in Nasopharyngeal Carcinoma. Cancer Management and Research, 12, 11585-11596.

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Multiplex IHC Analysis of PD-L1, CD8, and CD163

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IHC for PD-L1, (catalog no. 13684; clone E1L3N, rabbit mcl., dilution 1:400, Cell Signaling Technology), CD8 (NCL-L-CD8-4B11; mouse mcl., dilution 1:400, Leica), and CD163 (NCL-L-CD163; mouse mcl., dilution 1:500, Leica) was performed on automated BondRx platform (Leica Biosystems) as per standard IHC protocol. PD-L1 staining was called positive if it is expressed on >1% tumor cells or immune cells. CD8- and CD163-positive immune cells were counted on Halo image analysis platform (Halo 2.3; Indica Laboratories) per tissue area (μm²).

Sridharan V., Neyaz A., Chogule A., Baiev I., Reyes S., Barr Fritcher E.G., Lennerz J.K., Sukov W., Kipp B., Ting D.T., Deshpande V, & Goyal L. (2022). FGFR mRNA Expression in Cholangiocarcinoma and Its Correlation with FGFR2 Fusion Status and Immune Signatures. Clinical Cancer Research, 28(24), 5431-5439.

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Immunohistochemical Analysis of PD-L1, CD68, and CD163

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TMA slides were dewaxed in xylene and ethanol and steamed in a 2100 Retriever (Aptum Bioologics, Southampton, UK) with pH 6.0 sodium citrate buffer to retrieve antigen epitopes. Once the samples had cooled, endogenous peroxidase activity was blocked using 3% hydrogen peroxide, and biotin was blocked using a biotin blocking kit. The slides were incubated at 4 °C overnight with a primary antibody: PD-L1 with an anti-human PD-L1 rabbit monoclonal antibody (dilution 1:100; clone SP142; Spring Bioscience, Pleasanton, CA, USA), CD68 with an anti-human CD68 mouse monoclonal antibody (dilution 1:200; KP-1; Agilent, Santa Clara, CA, USA), or CD163 with an antihuman CD163 mouse monoclonal antibody (dilution 1:200; NCL-L-CD163; Leica, Nussloch, Germany). The secondary antibody was a ready-to-use VECTASTAIN ABC HRP Kit (Vector Laboratories, Burlingame, CA, USA).

Harada K., Dong X., Estrella J.S., Correa A.M., Xu Y., Hofstetter W.L., Sudo K., Onodera H., Suzuki K., Suzuki A., Johnson R.L., Wang Z., Song S, & Ajani J.A. (2018). Tumor-associated macrophage infiltration is highly associated with PD-L1 expression in gastric adenocarcinoma. Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association, 21(1).

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