Ab85792

Manufactured by Abcam

Sourced in United States

Ab85792 is a primary antibody produced in rabbit. It is designed for use in immunocytochemistry and western blotting applications.

Automatically generated - may contain errors

Lab products found in correlation

Ab133616, by Abcam (3 mentions) Ab959, by Abcam (2 mentions) Opal multiplex kit, by PerkinElmer (1 mentions) Prolong diamond antifade mountant containing dapi, by Thermo Fisher Scientific (1 mentions) Ab27988, by Abcam (1 mentions) Ab8021, by Abcam (1 mentions) Ab150440, by Abcam (1 mentions) Ab20181, by Abcam (1 mentions) Ab27969, by Abcam (1 mentions) Anti cd31, by Cell Signaling Technology (1 mentions) Dfc310 fx microscope, by Leica (1 mentions) Ab41532, by Abcam (1 mentions) Ab28244, by Abcam (1 mentions) Ab32575, by Abcam (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Enhanced chemiluminescence ecl reagent, by Thermo Fisher Scientific (1 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (1 mentions) Pentobarbital sodium, by Merck Group (1 mentions) Matrigel, by BD (1 mentions) Facs lysing solution, by BD (1 mentions)

5 protocols using ab85792

Multiplex Immunofluorescence Profiling of Tissue Samples

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Tissue samples were fixed in formalin, embedded in paraffin, and sectioned. These specimens were incubated with the following antibodies: anti-CD3 (A045229–2; DAKO), anti-CD4 (ab133616; Abcam), anti-Pan-CK (ab27988; Abcam), anti-CD31 (3528; Cell Signaling Technology), anti-SLAMF7 (HPA055945; Sigma-Aldrich), anti-CX3CR1 (ab8021; Abcam), anti-T-bet (ab150440; Abcam), anti-GATA3 (MA1028; Invitrogen), anti-Ror gamma (ab212496; Abcam), anti-CXCR5 (clone: MAB190; R&D Systems), anti-FoxP3 (clone: 98377; Cell Signaling Technology), anti-CD8 (ab85792; Abcam), anti-PD1 (B13300; Lifespan Bioscience), anti-GZMB (ab4095; Abcam), anti-HLA-DR (ab20181; Abcam), anti-TGF-β (ab27969; Abcam) and anti-cleaved caspase-3 (9664; Cell Signaling Technology) followed by incubation with a secondary antibody using an OpalTM Multiplex Kit (Perkin Elmer). The samples were mounted with ProLongTM Diamond Antifade mountant containing DAPI (Invitrogen).

Kaneko N., Boucau J., Kuo H.H., Perugino C., Mahajan V.S., Farmer J.R., Liu H., Diefenbach T.J., Piechocka-Trocha A., Lefteri K., Waring M.T., Premo K.R., Walker B.D., Li J.Z., Gaiha G., Yu X.G., Lichterfeld M., Padera RF J.r, & Pillai S. (2022). Temporal changes in T cell subsets and expansion of cytotoxic CD4+ T cells in the lungs in severe COVID-19. Clinical Immunology (Orlando, Fla.), 237, 108991.

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Immunohistochemical Profiling of Tumor Microenvironment

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The cancer samples were labeled for PS1 (NB100-74510, Novus Biologicals, Centennial, CO, USA; PA5-13214, Invitrogen), FAP (ab28244, Abcam), α-SMA (ab32575, Abcam), fibroblast-specific protein (FSP) (ab41532, Abcam), and anti-CD8 (ab85792, Abcam) by immunohistochemistry. Primary antibodies were added to each section. The sections were covered with 4plus biotinylated goat antirabbit IgG (Biocare Medical, Pacheco, CA, USA) and incubated in a humidified chamber for 30 min at room temperature. Primary tumors excised from mouse xenografts were snap frozen for subsequent histological examination. Images were collected using a Leica DFC310 FX microscope (Buffalo Grove, IL, USA). The stained sections were evaluated for the number of CD8+ positive cells using anti-CD8a antibody (550,298, BD Biosciences, Franklin Lakes, NJ, USA). Scoring was divided into the following categories: <5 cells/high-power field (HPF; 40 × magnification), 5–10 cells/HPF, 10–15 cells/HPF, 15–30 cells/HPF, and >30 cells/HPF. The topographic distribution of positive cells was also evaluated. Location relative to the neoplastic follicle (perifollicular, intrafollicular, or neither, i.e., evenly distributed) was recorded.

Zhang H., Jiang R., Zhou J., Wang J., Xu Y., Zhang H., Gu Y., Fu F., Shen Y., Zhang G., Feng L., Zhang X., Chen Y, & Shen F. (2020). CTL Attenuation Regulated by PS1 in Cancer-Associated Fibroblast. Frontiers in Immunology, 11, 999.

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Immunohistochemical Characterization of T-cell Subsets

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Matrigel and FACS lysing solution were purchased from BD Biosciences (San Jose, CA, USA). Hydrogen peroxide (H₂O₂), 3,3′-diaminobenzidine and pentobarbital sodium were purchased from Sigma-Aldrich (St. Louis, MO, USA). RIPA lysis buffer and enhanced chemiluminescence (ECL) reagent were purchased from Pierce Biotechnology, Inc. (Rockford, IL, USA). Rabbit polyclonal antibody against PC5-CD3 was purchased from Beckman Coulter, Inc. (Brea, CA, USA). Rabbit monoclonal antibody against CD3 (ab109531), and rabbit polyclonal antibodies against CD4 (ab70951), CD8 (ab85792) and FoxP3 (ab10563) were purchased from Abcam (Cambridge, MA, USA). Rabbit polyclonal antibody against GAPDH was purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Horseradish peroxidase (HRP)-coupled goat polyclonal anti-rabbit secondary IgG (sc-2004) and biotinylated goat polyclonal anti-rabbit IgG (sc-2040) secondary antibodies, as well as the avidin-biotin-HRP complex were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). RPMI-1640 medium and fetal bovine serum (FBS) were purchased from Life Technologies (Grand Island, NY, USA).

LIU X., LI H., LIU J., GUAN Y., HUANG L., TANG H, & HE J. (2014). Immune reconstitution from peripheral blood mononuclear cells inhibits lung carcinoma growth in NOD/SCID mice. Oncology Letters, 8(4), 1638-1644.

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Immunohistochemical Quantification of Immune Cells in CRC

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Four serial sections of 5 μm per paraffin block are obtained for the following immunohistochemistry staining. These sections were first baked at 60 °C and then deparaffinized in xylene and ethanol. After hydration, 3% hydrogen peroxidase was utilized to block endogenous peroxidase activity. Standard antigen retrieval was conducted via heating the sections immersed in citric acid solution (pH = 6.0) in a pressure boiler. Subsequently, these slides were incubated with the primary antibodies [CD20 (60271-1-Ig, Proteintech, 1:5000); CD4(ab133616, Abcam, 1:500), CD8(ab85792, abcam, 1:400), CD68(ab959, Abcam, 1:6000)] at 4°Covernight, and then incubated with second antibody. After 3,3'-diaminobenzidine tetrahydrochloride staining and hematoxylin counterstaining, the slides were scanned for further quantitative analysis. The density of CD4 + , CD20,CD68 and CD8 + T cells both invasive margin (IM) and in the core of the tumor (CT) were automatically calculated using ImageJ software (version 1.48). The software generally contains positive cells and a positive nucleus, and we used its ratio (positive cells/positive nucleus) to represent the expression status of four immune cells in CRC tissues.

Cao Y., Zheng X., Hu Y., Li J., Huang B., Zhao N., Liu T., Cai K, & Tian S. (2023). Levels of systemic inflammation response index are correlated with tumor-associated bacteria in colorectal cancer. Cell Death & Disease, 14(1), 69.

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Immunohistochemical Quantification of Immune Cell Markers

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Four serial sections of 5 μm per paraffin block for subsequent immunohistochemistry staining were obtained. Initially, these sections were subjected to baking at 60 °C, followed by deparaffinization in xylene and ethanol. After hydration, we used 3 per cent hydrogen peroxidase to block endogenous peroxidase activity. For standard antigen retrieval, the sections were heated in a citric acid solution (pH = 6.0) using a pressure boiler. Subsequently, the slides were incubated with primary antibodies [CD4 (ab133616, Abcam, 1:500); CD8 (ab85792, Abcam, 1:400); CD20 (60271-1-Ig, Proteintech, 1:5000); CD68 (ab959, Abcam, 1:6000)] overnight at 4 °C, followed by incubation with a secondary antibody. After staining with 3,3ʹ-diaminobenzidine tetrahydrochloride and counterstaining with haematoxylin, the slides were scanned for further quantitative analysis. The density of CD4+, CD8+, CD20 and CD68 T cells at the invasive margin (IM) was automatically calculated using ImageJ software (version 1.48).

Cao Y., Shang F., Jin M., Deng S., Gu J., Mao F., Qin L., Wang J., Xue Y., Jiang Z., Cheng D., Liu L., Nie X., Liu T., Liu H, & Cai K. (2023). Changes in Bacteroides and the microbiota in patients with obstructed colorectal cancer: retrospective cohort study. BJS Open, 7(6), zrad105.

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