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Cd68 clone pg m1

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CD68 (clone PG-M1) is a monoclonal antibody used for the identification and quantification of macrophages and monocytes in a variety of tissues and cell preparations. It recognizes a 110-kDa glycoprotein, a member of the lysosome-associated membrane protein (LAMP) family, which is highly expressed in human monocytes and macrophages.

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Lab products found in correlation

Clone emr8 5, by Abcam (1 mentions) Cd11c clone ep1347y, by Abcam (1 mentions) Rnalater, by Thermo Fisher Scientific (1 mentions) Bx40 microscope, by Olympus (1 mentions) Ultravision detection system, by Thermo Fisher Scientific (1 mentions) Citrate buffer, by BioGenex (1 mentions) Automated link 48 platform, by Agilent Technologies (1 mentions) Cd8 clone 144b, by Abcam (1 mentions) Tween 20, by Thermo Fisher Scientific (1 mentions) Cd20 clone l26, by Roche (1 mentions) Cd4 clone ep204, by Cell Marque (1 mentions) Cd8 clone sp57, by Roche (1 mentions) Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)

4 protocols using cd68 clone pg m1

1

Immunohistochemical Analysis of Tumor-Infiltrating Immune Cells

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ITM biopsies from Pt#5 obtained before and after VACCIMEL immunization, and following nivolumab treatment, were formalin-fixed and paraffin embedded for histological analyses. From the latter ITM, a fresh punch biopsy was preserved at −80°C in RNAlater (Ambion) for RNA-seq analysis. Five-micrometer tumor sections were stained with anti-human monoclonal Abs to HLA- class I (A, B, C) (clone EMR8-5, Abcam), PMEL (clone hmb45, Dako), TYR (clone T311, Santa Cruz), CD8 (cloneC8/144, Dako), CD68 (clone PG-M1, Abcam), CD11c (clone EP1347Y, Abcam), DC-LAMP (clone 1010E1.01, Dendritics), and CD163 (clone 10D6, Invitrogen). The Avidin-Biotin-Peroxidase (ABC) system (Vectastain, Vector Labs) was used. Sections were examined by optical microscopy (Olympus BX40 microscope, DP2-BSW software). Quantification of the tumor-infiltrating immune populations in the different ITM biopsies was performed on the 1-mm2 hot spot area by duplicate (ImageJ software), distinguishing zones with peritumoral (PT) and intratumoral (IT) infiltrate. Immune cell counts in the different ITM biopsies were compared by Tukey’s multiple comparisons test; p < 0.05 was considered as statistically significant.
Mordoh A., Aris M., Carri I., Bravo A.I., Podaza E., Pardo J.C., Cueto G.R., Barrio M.M, & Mordoh J. (2022). An Update of Cutaneous Melanoma Patients Treated in Adjuvancy With the Allogeneic Melanoma Vaccine VACCIMEL and Presentation of a Selected Case Report With In-Transit Metastases. Frontiers in Immunology, 13, 842555.
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2

Immune Profiling of Prostate Tumors

Cited 1 time
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Pre- and post-chemotherapy plasma vascular endothelial growth factor (VEGF) were measured. Tumor expression of tissue VEGF was characterized by tissue microarray. Methods for these assays have been described [20 (link)]. Also, serum total testosterone levels were obtained prior to and after neoadjuvant chemotherapy.
Immune infiltration was examined via immunohistochemistry with a panel of 8 markers in primary prostatic tumors and adjacent normal tissue using a tissue microarray (TMA) of 50 patients treated neoadjuvantly with docetaxel and mitoxantrone. Heat mediated antigen retrieval was performed with citrate buffer (BioGenex, Fremont, CA), with the exception of retrieval prior to CD4 staining, which was achieved using 10 mM Tris/1 mM EDTA. Staining was performed with the Ultravision Detection System (Thermo Scientific, Waltham, MA), followed by counterstaining with methyl green. Antibodies CD3, CD4, CD8, CD15, and CD163 were from Thermo Scientific, CD68 (clone PG-M1) and CD20 were from AbCAM (Cambridge, UK), and FoxP3 was from eBioscience (San Diego, CA). Slides were digitally scanned with an Aperio ScanScope CT Slide Scanner at 40x objective and individual TMA cores were analyzed separately using TMALab (Aperio, Buffalo Grove, IL) and the nuclear stain detection algorithm to quantify the number of positive cells within each core. Data is displayed as cell density (cells/mm2).
Bergstrom C.P., Ruffell B., Ho C.M., Higano C.S., Ellis W.J., Garzotto M., Beer T.M, & Graff J.N. (2017). Docetaxel and Mitoxantrone Prior to Radical Prostatectomy in Men with High Risk Prostate Cancer: Ten-Year Follow Up and Immune Correlates. Anti-cancer drugs, 28(1), 120-126.
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3

Tumor Immune Microenvironment Profiling

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We obtained tumor tissues from different origins, including the lung, liver, lymph nodes, soft tissue and bones. According to manufacturer’s recommendations, IHC was conducted using 4–5 μm formalin-fixed and paraffin-embedded (FFPE) sections. The PD-L1 clone 28-8 pharmDx kit and the Dako Automated Link 48 platform were used to measure the expression of PD-L1. CD4 (clone B468A, diluted at 1:200, Santa Cruz, Texas, USA) and CD8 (clone 144B, diluted at 1:100, Abcam, Cambridge, UK) expression in T cells, and CD68 (clone PG-M1, diluted at 1:600, Abcam, Cambridge, UK) expression on macrophages were also assessed. Two pathologists independently scored the stained tissues, and the clinical parameters were kept confidential from these two pathologists.
The tumor proportion score (TPS) was used to assess PD-L1 expression. The percentage of tumor cells stained with partial or complete membranes was the definition of TPS. The positive expression of PD-L1 was considered as TPS ≥ 1%. Among all nucleated cells in the tumor mesenchyme, the proportion of positive cells for CD4, CD8, and CD68 expression was assessed. Positivity on lymphocytes was set at 5, 5, and 20% for CD4, CD8, and CD68, respectively. Based on PD-L1 expression and CD8+ T cell infiltration, the tumor microenvironment was divided into three subgroups (both negative, both positive, or single-positive).
Xie M., Li N., Xu X., Xu Y., Li H., Zhu L., Sheng J., Zhou Z, & Fan Y. (2022). The Efficacy of PD-1/PD-L1 Inhibitors in Patients with Liver Metastasis of Non-Small Cell Lung Cancer: A Real-World Study. Cancers, 14(17), 4333.
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4

Multiplex Immunofluorescence Staining Protocol

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10x Tris buffered saline, Tween-20, and ProLong Gold antifade mountant with DAPI were from ThermoFisher Scientific (Waltham, MA). The Manual Opal 7-color IHC Kit was obtained from Akoya Biosciences (Marlborough, MA). The following primary antibodies were used for immunofluorescence: CD4 (clone EP204, CellMarque, 1:1250 dilution, antigen retrieval in AR9 buffer), CD8 (clone SP57, Ventana, undiluted, antigen retrieval in AR9 buffer), CD20 (clone L26, Ventana, 1:2 dilution, antigen retrieval in AR6 buffer), CD68 (clone PG-M1, Abcam, 1:250 dilution, antigen retrieval in AR9 buffer), CD138 (clone B-A38, Ventana, 1:2 dilution, antigen retrieval in AR6 buffer, pan-CK (PA1–27114, Invitrogen, antigen retrieval in AR9 buffer, 1:500 dilution), AF750 goat anti-rabbit Ig (clone Ab175733, Abcam, 1:200 dilution).
D’Souza I., Poorkaj P., Hong M., Nochlin D., Lee V.M., Bird T.D, & Schellenberg G.D. (1999). Missense and silent tau gene mutations cause frontotemporal dementia with parkinsonism-chromosome 17 type, by affecting multiple alternative RNA splicing regulatory elements. Proceedings of the National Academy of Sciences of the United States of America, 96(10), 5598-5603.
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