Cd8 clone c8 144b

Manufactured by Agilent Technologies

Sourced in United States, United Kingdom, Denmark, France

The CD8 (clone C8/144B) is a laboratory reagent used for the detection and analysis of CD8-positive cells in flow cytometry applications. It is a monoclonal antibody that specifically binds to the CD8 surface antigen, which is expressed on cytotoxic T cells and a subset of natural killer cells. This reagent can be used to identify and quantify CD8-positive cells in various biological samples.

Automatically generated - may contain errors

Lab products found in correlation

38 protocols using cd8 clone c8 144b

Tumor Immune Cell Infiltration Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Intratumoral immune cell infiltration at different time points was determined by immunohistochemistry assays of FFPE tumor blocks (3 µm). Immunohistochemistry of the FFPE tumor samples was performed on a BenchMark Ultra autostainer. Briefly, paraffin sections were cut at 3 µm, heated at 75°C for 28 min and deparaffinized in the instrument with EZ prep solution (Ventana Medical Systems). Heat-induced antigen retrieval was carried out using Cell Conditioning 1 (CC1, Ventana Medical Systems) for 32 min at 95°C (CD3, CD8), 48 min at 95°C (PD-L1).
CD3 was detected using clone SP7 (1:100 dilution, 32 min at 37°C, Spring/ITK), CD8 clone C8/144B (1:200 dilution, 32 min at 37°C, DAKO/Agilent) and PD-L1 using clone 22C3 (1:40 dilution, 1-hour room temperature, DAKO/Agilent). Bound antibody was detected using the OptiView DAB Detection Kit (Ventana Medical Systems). Slides were counterstained with Hematoxylin and Bluing Reagent (Ventana Medical Systems). Scoring was performed by a blinded pathologist.

Rozeman E.A., Versluis J.M., Sikorska K., Hoefsmit E.P., Dimitriadis P., Rao D., Lacroix R., Grijpink-Ongering L.G., Lopez-Yurda M., Heeres B.C., van de Wiel B.A., Flohil C., Sari A., Heijmink S.W., van den Broek D., Broeks A., de Groot J.W., Vollebergh M.A., Wilgenhof S., van Thienen J.V., Haanen J.B, & Blank C.U. (2023). IMPemBra: a phase 2 study comparing pembrolizumab with intermittent/short-term dual MAPK pathway inhibition plus pembrolizumab in patients with melanoma harboring the BRAFV600 mutation. Journal for Immunotherapy of Cancer, 11(7), e006821.

+ Open protocol

+ Expand

Multiplex Immunofluorescence Profiling of PDAC

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human PDAC samples (n = 6) were fixed in 4% paraformaldehyde and embedded in paraffin. Five-micrometer sections were used for the immunofluorescence staining. Immunofluorescence staining on FFPE tissue was performed using the Opal 7-Color Automated Immunohistochemistry Kit (cat: NEL821001KT, Akoya Biosciences). A multiplex panel of immune markers was developed with antibodies against: CXCR4 (clone EPUMBR3, cat: ab181020, dilution 1:300, Abcam), CD8 (clone C8/144B, cat: M710301–2, dilution 1:200, Dako/Agilent), CD68 (clone PG-M1, cat: M087601–2, dilution 1:400, Dako/Agilent), Cytokeratin (Cytokeratin 7, clone OV-TL, cat: M701801–2, dilution 1:500, Dako/Agilent + Cytokeratin 19, clone A53-B/A2.26, cat: 760–4281, RUI, Cell Marque/Roche). The staining procedure was performed using an automated staining system (BOND-RX; Leica Biosystems). All markers were sequentially applied and paired with respective Opal fluorophores. To visualize cell nuclei, the tissue was stained with 4‘,6-diamidino-2-phenylindole (spectral DAPI, Akoya Biosciences). Stained slides were scanned using Mantra 2 Quantitative Pathology Workstation (Akoya Biosciences) and representative images from each tissue were acquired with the Mantra Snap software version 1.0.4. Spectral unmixing, multispectral image acquisition was carried out using the InForm Tissue Analysis Software version 2.4.10 (Akoya Biosciences).

Kocher F., Puccini A., Untergasser G., Martowicz A., Zimmer K., Pircher A., Baca Y., Xiu J., Haybaeck J., Tymoszuk P., Goldberg R.M., Petrillo A., Shields A.F., Salem M.E., Marshall J.L., Hall M., Korn W.M., Nabhan C., Battaglin F., Lenz H.J., Lou E., Choo S.P., Toh C.K., Gasteiger S., Pichler R., Wolf D, & Seeber A. (2022). Multi-omic Characterization of Pancreatic Ductal Adenocarcinoma Relates CXCR4 mRNA Expression Levels to Potential Clinical Targets. Clinical Cancer Research, 28(22), 4957-4967.

+ Open protocol

+ Expand

Multiplex Immunofluorescence of Renal Cell Carcinoma

Check if the same lab product or an alternative is used in the 5 most similar protocols

In-House RCC samples (n = 4) after surgery were fixed in 4% paraformaldehyde, embedded in paraffin and five-micrometer sections were used for the immunofluorescence staining. Multiplex IHC was performed using Opal 6-plex Detection Kit (cat: NEL821001KT, Akoya Biosciences, Menlo Park, USA). A multiplex panel of immune markers was developed with antibodies against CXCR3 (clone EPR25373-32, cat: ab288437, dilution 1:200, Abcam, Cambridge, MA, USA), CD4 (clone EP204, cat: 104R-26, dilution 1:50, Cell Marque), CD8 (clone C8/144B, cat: M710301-2, dilution 1:200, Dako/Agilent, Santa Clara, CA, USA), CD68 (clone PG-M1, cat: M087601-2, dilution 1:250, Dako/Agilent), cytokeratin (clone AE1/AE3, cat: MA5-13156, dilution 1:500, Thermo-Fisher). The staining procedure was performed using an automated staining system (BOND-RX; Leica Biosystems, Vienna, Austria). To visualize cell nuclei, the tissue was stained with 4’,6-diamidino-2-phenylindole (spectral DAPI, Akoya Biosciences). Slides were scanned at 20x magnification using Mantra 2 Quantitative Pathology Workstation (Akoya Biosciences) and representative images from each tissue were acquired with the Mantra Snap software version 1.0.4. Image spectral deconvolution, multispectral image analysis and cell phenotyping was carried out using the InForm Tissue Analysis Software version 2.4.10 (Akoya Biosciences).

Pichler R., Siska P.J., Tymoszuk P., Martowicz A., Untergasser G., Mayr R., Weber F., Seeber A., Kocher F., Barth D.A., Pichler M, & Thurnher M. (2023). A chemokine network of T cell exhaustion and metabolic reprogramming in renal cell carcinoma. Frontiers in Immunology, 14, 1095195.

+ Open protocol

+ Expand

Multiplex Immunostaining of TFE3, HIF1A, CD31, CD8, and PD-L1

Check if the same lab product or an alternative is used in the 5 most similar protocols

IHC and multiple immunofluorescence were performed as previously described^{32 (link)}. Commercially available primary anti-TFE3 (clone MRQ-37, 1:100, MXB biotechnologies, Fujian, China), anti-HIF1A (1:5000, Novus Biologicals, Colorado, USA), anti-CD31 (clone UMAB30, ready to use, ZSGB-BIO, Beijing, ChinaMXB), CD8 (clone C8/144B, ready to use, Dako, Copenhagen, DEN) and PD-L1 (clone 22C3, 1:50, Dako) were used in this study. Multiplex immunofluorescence staining was performed with a PANO 7-plex kit (0004100100, Panovue, Beijing, CHN). CD8, macrophage, and NK cells expression were quantified as positive cell density (cell number per mm²). PD-L1 expression was assessed by tumor proportion score, which was defined as the percentage of tumor cells with membranous PD-L1 staining. PD-L1 expression >1% was defined as positivity.

Sun G., Chen J., Liang J., Yin X., Zhang M., Yao J., He N., Armstrong C.M., Zheng L., Zhang X., Zhu S., Sun X., Yang X., Zhao W., Liao B., Pan X., Nie L., Yang L., Chen Y., Zhao J., Zhang H., Dai J., Shen Y., Liu J., Huang R., Liu J., Wang Z., Ni Y., Wei Q., Li X., Zhou Q., Huang H., Liu Z., Shen P., Chen N, & Zeng H. (2021). Integrated exome and RNA sequencing of TFE3-translocation renal cell carcinoma. Nature Communications, 12, 5262.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Tumor Microenvironment

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemistry was performed to detect the presence of PD-L1, CD8, FoxP3, and 4-1BB/CD137 in the whole tumor and invasive margin (IM) of pretreatment tumor biopsies. Immunohistochemistry testing of PD-L1 (clone E1L3N; Cell Signaling, Danvers, MA), CD8 (clone C8/144B; Dako, Carpinteria, CA), FoxP3 (clone 236A/E7; Cell Signaling), and 4-1BB/CD137 (BBK-2; ThermoFisher, Rockford, IL) was performed by Mosaic Laboratories, LLC (Lake Forest, CA).

Cohen E.E., Pishvaian M.J., Shepard D.R., Wang D., Weiss J., Johnson M.L., Chung C.H., Chen Y., Huang B., Davis C.B., Toffalorio F., Thall A, & Powell S.F. (2019). A phase Ib study of utomilumab (PF-05082566) in combination with mogamulizumab in patients with advanced solid tumors. Journal for Immunotherapy of Cancer, 7, 342.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Tissue Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bone marrow biopsies were decalcified in EDTA prior to paraffin embedding. 3 μm sections were cut from formalin-fixed, paraffin-embedded samples. Upper gastrointestinal biopsies were routinely stained with hematoxylin and eosin stain and periodic acid–Schiff stain. Bone marrow biopsies were routinely stained with naphthol AS-D chloroacetate esterase (NASDCL). Immunohistochemical staining was performed using a horseradish peroxidase catalyzed brown chromogen reaction together with ready-to-use antibodies in an automated staining system (Dako Autostainer Link®; Dako, Glostrup, Denmark), following the manufacturer's guidelines. Depending on the tissue section size, up to three droplet zones are stained with 100 μl antibody solution per droplet zone. The following antibodies were used: CD3 (rabbit polyclonal, Dako), CD4 (clone 4B12, Dako), CD8 (clone C8/144B, Dako), CD19 (clone LE-CD19, Dako), CD20 (clone L26, Dako), CD38 (clone SPC 32, Novocastra, Newcastle upon Tyne, UK), IgM, IgG, IgA (rabbit polyclonal antibodies; Dako). Photos were taken on an Olympus BX51 microscope (Olympus Germany, Hamburg, Germany) with the AxioCam MRc camera (Zeiss, Jena, Germany).

Schubert D., Bode C., Kenefeck R., Hou T.Z., Wing J.B., Kennedy A., Bulashevska A., Petersen B.S., Schäffer A.A., Grüning B.A., Unger S., Frede N., Baumann U., Witte T., Schmidt R.E., Dueckers G., Niehues T., Seneviratne S., Kanariou M., Speckmann C., Ehl S., Rensing-Ehl A., Warnatz K., Rakhmanov M., Thimme R., Hasselblatt P., Emmerich F., Cathomen T., Backofen R., Fisch P., Seidl M., May A., Schmitt-Graeff A., Ikemizu S., Salzer U., Franke A., Sakaguchi S., Walker L.S., Sansom D.M, & Grimbacher B. (2014). Autosomal-dominant immune dysregulation syndrome in humans with CTLA4 mutations. Nature medicine, 20(12), 1410-1416.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Gliosis and Lymphocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Resected tissues were fixed in 10% buffered formalin and embedded in paraffin. In each case, a search for gliosis and lymphocytes in the serial cryosections was performed using both a routine hematoxylin and eosin (H&E) stain and immunohistochemical reactions with anti-GFAP (clone 6F2, Dako, diluted 1:1000) and anti-CD45 (leukocyte common antigen - LCA, clone 2B11 + PD7/26, Dako, diluted 1:100) antibodies, respectively. Provided that lymphocytes were found, the contiguous sections were subjected to immunohistochemistry to establish their phenotype using antibodies against the lymphocytic surface antigens CD20 (clone L26, Dako, diluted 1:300), CD8 (clone C8/144B, Dako, diluted 1:200) and CD4 (clone 4B12, Novocastra, diluted 1:20). The antigen-antibody complexes were visualized with biotin-streptavidin detection systems (N-Histofine Simple Stain MAX PO, Nichirei Corporation, Tokyo, Japan); chromogenic development was performed using 3,3-diaminobenzidine. All sections were counterstained slightly with Harris’ hematoxylin. Positive and negative controls were used in each assay.

Liba Z., Vaskova M., Zamecnik J., Kayserova J., Nohejlova H., Ebel M., Sanda J., Ramos-Rivera G.A., Brozova K., Liby P., Tichy M, & Krsek P. (2020). An immunotherapy effect analysis in Rasmussen encephalitis. BMC Neurology, 20, 359.

+ Open protocol

+ Expand

Immunohistochemical Staining for Mismatch Repair Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemical staining was performed using the BenchMark XT autostainer (Ventana Medical Systems, Tucson, AZ, USA) with an UltraView detection kit (Ventana Medical Systems, Basel, Switzerland). Commercially-available antibodies were used for immunohistochemical staining; CD8 (Clone C8/144B, ready to use; Dako, Carpinteria, CA, USA), hMLH1 (Clone M1, ready to use; Ventana, Basel, Switzerland), hMLH2 (Clone G219-1,129, 1:100 dilution; Abnova, Taipei City, Taiwan), hMLH6 (Clone 44, 1:100 dilution; Cell Marque, Rocklin, CA, USA), and PMS2 (Clone A16-4, ready to use; Ventana Medical Systems, Basel, Switzerland). The normal lymph node was used as positive control of CD8 stain. For MLH1, MLH2, MLH6, and PMS2, inflammatory cells and stromal tissue were used as internal positive control.

Ahn S., Lee J.C., Shin D.W., Kim J, & Hwang J.H. (2020). High PD-L1 expression is associated with therapeutic response to pembrolizumab in patients with advanced biliary tract cancer. Scientific Reports, 10, 12348.

+ Open protocol

+ Expand

Immunohistochemical Staining of MMR and PD-L1 Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemical staining was performed using the Envision™ FLEX (DAKO, Glostrup, Denmark) and a Dako Autostainer Plus (DAKO) according to manufacturer’s instructions. Antibodies used on whole slides included anti-MLH1 (G168-15, dilution 1:100), anti-PMS2 (A16-4, 1:500), and anti-MSH6 (clone 44, 1:200) from BD Bioscience (San Jose, CA, USA), anti-MSH2 (25D12, 1:200) from Novocastra (Newcastle, UK), and PD-L1 (clone Sp142, diluted 1:100, Spring Bioscience, Ventana), while tissue microarray (TMA) (containing 2 × 1 mm biopsies from each case) immunostaining was performed using antibodies against CD3 (A0452, diluted 1:100, DAKO), CD8 (clone C8/144B, diluted 1:100, DAKO), Beta-2-Microglobulin (A0072, No pretreatment, dilution 1:600, DAKO), and CD68 (clone PG-M1 dilution 1:100). Control tissues such as brain, normal colon tissue and tonsils were used where appropriate. A pilot study of 46 tumors investigating the correlation between whole slide and tissue microarray scorings found a high correlation between the two different methods (Pearson correlation ranged from 0.75 to 0.82).

Walkowska J., Kallemose T., Jönsson G., Jönsson M., Andersen O., Andersen M.H., Svane I.M., Langkilde A., Nilbert M, & Therkildsen C. (2018). Immunoprofiles of colorectal cancer from Lynch syndrome. Oncoimmunology, 8(1), e1515612.

+ Open protocol

+ Expand

Liver Biopsy Immunohistochemical Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Liver biopsies were obtained for pathological examination. Standard morphological evaluation was based on hematoxylin and eosin (H&E) sections of formalin fixed specimens. To characterize the inflammatory infiltrates, immunohistochemical markers for macrophages (CD68, clone PG-M1, Dako, Glostrup, Denmark; CD163, clone 10D6, Thermo Scientific, Fremont, CA, USA), T lymphocytes (CD8, clone C8/144B, Dako, Glostrup, Denmark) and erythroid cells (Glycophorin, clone JC159, Dako, Glostrup, Denmark) were applied. Four-micron thick tissue sections were deparaffinised with xylene and rehydrated with graded alcohols. After washing in distilled water, sections were immersed in 3% hydrogen peroxide to block endogenous peroxidase.

Prencipe G., Bracaglia C., Caiello I., Pascarella A., Francalanci P., Pardeo M., Meneghel A., Martini G., Rossi M.N., Insalaco A., Marucci G., Nobili V., Spada M., Zulian F, & De Benedetti F. (2019). The interferon-gamma pathway is selectively up-regulated in the liver of patients with secondary hemophagocytic lymphohistiocytosis. PLoS ONE, 14(12), e0226043.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!