Rabbit anti human cd8

Manufactured by Abcam

Sourced in United States

Rabbit anti-human CD8 is a primary antibody that binds to the CD8 protein, which is expressed on the surface of certain T cells. It can be used in various immunological applications to detect and study CD8-positive cells.

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Lab products found in correlation

Bovine serum albumin (bsa), by Jackson ImmunoResearch (1 mentions) Normal goat serum (ngs), by Jackson ImmunoResearch (1 mentions) Diva decloaking buffer, by Biocare Medical (1 mentions) Hoechst dye, by Thermo Fisher Scientific (1 mentions) Alexa fluor 568 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 goat anti rat, by Thermo Fisher Scientific (1 mentions) Clone cd3 12, by Abcam (1 mentions) Prolong gold, by Thermo Fisher Scientific (1 mentions) Horseradish peroxidase conjugated anti rabbit igg, by Cell Signaling Technology (1 mentions) Anti mouse foxp3 antibody, by Thermo Fisher Scientific (1 mentions) Tim 3, by BioLegend (1 mentions) Tcs sp8 confocal system, by Leica (1 mentions) Alexa fluor 488 goat anti rabbit igg, by Abcam (1 mentions) Goat anti rabbit igg cy3, by Abcam (1 mentions) X image analysis software, by Leica (1 mentions) Anti human il 4 apc, by BioLegend (1 mentions) Ficoll paque plus, by GE Healthcare (1 mentions) Aim 5 medium, by Thermo Fisher Scientific (1 mentions) Facs lysing solution, by BD (1 mentions) Mouse igg2a, by Thermo Fisher Scientific (1 mentions)

6 protocols using rabbit anti human cd8

Multiplex Immunofluorescence Staining of FFPE Tumor Sections

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Formalin-fixed paraffin-embedded tumor sections were deparaffinized and pretreated with DIVA decloaking buffer (Biocare medical) at 125°C. Sections were blocked for 1 hour in 0.2% Bovine Serum Albumin (BSA)+2% normal goat serum (Jackson Immuno). Sections were stained with rat anti-human CD3 (BioRad, clone CD3-12, #MCA1477) at 1:100 and rabbit anti-human CD8 (Abcam, polyclonal,#ab4055) diluted 1:100 in blocking buffer for 1 hour, 25°C. Sections were stained with goat anti-rat AlexaFluor 488 (Thermo, #A11006) and goat anti-rabbit AlexaFluor 568 (Thermo, #A11011) each diluted 1:500 in blocking buffer for 1 hour, 25°C. Sections were then stained for 10 min in Hoechst dye (Thermo, #H3570) at 2 μg/mL in ddH₂O, and coverslipped with Prolong Gold (Thermo #P36935). Sections were imaged at ×40 (Plan apo, NA 0.95) on a Nuance Multispectral camera (Akoya Biosciences) with a Nikon Eclipse Ci microscope and images were analyzed with InForm Tissue Analysis software, V.2.4 (Akoya Biosciences).

Gardell J.L., Matsumoto L.R., Chinn H., DeGolier K.R., Kreuser S.A., Prieskorn B., Balcaitis S., Davis A., Ellenbogen R.G, & Crane C.A. (2020). Human macrophages engineered to secrete a bispecific T cell engager support antigen-dependent T cell responses to glioblastoma. Journal for Immunotherapy of Cancer, 8(2), e001202.

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Multiparameter Immune Cell Profiling

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Alexa Fluor 488–conjugated anti-mouse CD45, anti-human CD45, violet 421–conjugated anti-mouse CD4 and CD8, anti-human CD4 and CD8, and phycoerythrin-conjugated anti-mouse or anti-human CD28, 41BB, NKG2D, CD39, PD-1, LAG3, TIM3, and FOXP3, as well as isotype control antibodies, were purchased from Biolegend (San Diego, CA). Rabbit anti-mouse and human CD3 and rabbit anti-human CD8 antibodies were purchased from Abcam (Cambridge, MA). Biotin anti-mouse NKG2D, mouse anti-human NKG2D, rabbit anti-human FOXP3 and anti-human Tbet antibodies were purchased from R&D Systems (Minneapolis, MN). Anti-mouse FOXP3 antibody was purchased from Thermo Fisher. Horseradish peroxidase–conjugated anti-rabbit IgG was purchased from Cell Signaling Technology (Danvers, MA).

Hu J., Sun C., Bernatchez C., Xia X., Hwu P., Dotti G, & Li S. (2018). T cell homing therapy for reducing regulatory T cells and preserving effector T cell function in large solid tumors. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(12), 2920-2934.

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Immunofluorescence Analysis of Colon Biopsies

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Paraffin-embedded sections of colonic biopsies were deparaffinized in xylene, rehydrated in decreasing concentrations of ethanol, and then heated in 0.01 M citric acid buffer (pH 6.0) for 15 min at 95°C to reveal antigens. After deparaffinization, sections were rinsed in blocking buffer (1× PBS/5% normal goat serum/0.3% Triton X-100) for 1 h at room temperature (RT) and then incubated sequentially with rabbit anti-human CD8 (1:100) (Abcam) overnight at 4°C, goat anti-rabbit-IgG-Cy3 (1:500) (Abcam) for 1 h at RT, rabbit anti-human cleaved (Asp175) caspase-3 (Cell Signaling Technology) overnight at 4°C, goat anti-rabbit-IgG-Alexa Fluor 488 (1:500) (Abcam) for 1 h at RT, and anti-human IL-4-APC (BioLegend) overnight at 4°C. The sections were mounted with a Vector Shield mounting solution containing 4′,6-diamidino-2-phenylindole (H-1200, Vector). Washing three times with 1× PBS was performed between incubations. Immunofluorescence images were acquired with a Leica TCS SP8 confocal system (Leica Microsystems) using a 20×/0.75 dry objective lens and analyzed with Leica X image analysis software (Leica). Post-acquisition processing (brightness, opacity, contrast, and color balance) was applied to the entire image and accurately reflects that of the original.

Zhang Y., Maksimovic J., Huang B., De Souza D.P., Naselli G., Chen H., Zhang L., Weng K., Liang H., Xu Y., Wentworth J.M., Huntington N.D., Oshlack A., Gong S., Kallies A., Vuillermin P., Yang M, & Harrison L.C. (2018). Cord Blood CD8+ T Cells Have a Natural Propensity to Express IL-4 in a Fatty Acid Metabolism and Caspase Activation-Dependent Manner. Frontiers in Immunology, 9, 879.

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Characterization of T cell responses

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The following were used: PGD₂ and LTE₄ (Enzo Life Science); TM30089 and montelukast (Cayman Chemical); rhIL-5, anti-CD3/28 antibodies, human CD8⁺ T cell isolation kit and anti-human CD294 or CD16 MicroBeads (Miltenyi Biotec Ltd); BD FACS Lysing Solution (BD Biosciences); X-VIVO-15 medium and Bronchial Epithelial Cell Growth Medium (Lonza); AIM V medium (Invitrogen); Ficoll-PaqueTM Plus (GE Healthcare); RNeasy Mini kit and Omniscript reverse transcription kit (Qiagen); Real time quantitative PCR Master Mix and probes (Roche); Primers (Eurofins MWG Operon); rhIL-6/SCF and anti-IL-4/5/13 neutralizing antibodies (Bio-techne); goat anti-human CD3 (Santa Cruz); rabbit anti-human CD8 (Abcam); peroxidase polymer-conjugated anti-rabbit and peroxidase polymer-conjugated anti-goat antibodies (Vector Laboratories); Fluorescein-tyramide and Cy5-tyramide (PerkinElmer); human myeloma IgE (Calbiochem); Anti-human GM-CSF antibody, Annexin V-APC and Human GM-CSF ELISA kit (BioLegend); IC fixation buffer, Permeabilisation Buffer, human IL-4/5/13 ELISA kits (eBioscience); and rhIL-2/4/GM-CSF (PeproTech) and other chemicals (Sigma-Aldrich).

Hilvering B., Hinks T., Stöger L., Marchi E., Salimi M., Shrimanker R., Liu W., Chen W., Luo J., Go S., Powell T., Cane J., Thulborn S., Kurioka A., Leng T., Matthews J., Connolly C., Borg C., Bafadhel M., Willberg C., Ramasamy A., Djukanović R., Ogg G., Pavord I., Klenerman P, & Xue L. (2018). Synergistic activation of pro-inflammatory type-2 CD8+ T lymphocytes by lipid mediators in severe eosinophilic asthma. Mucosal immunology, 11(5), 1408-1419.

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Immunophenotyping of VZV Infection

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The following primary antibodies and dilutions were used: mouse anti-human CD3 (Novocastra, Australia) (20 μg/mL), goat anti-human CD4 (R&D Systems, United States) (10 μg/mL), Rabbit anti-human CD8 (Abcam, United States) (2 μg/mL), mouse anti-human CD20 (Novocastra, Australia) (18 μg/mL), mouse anti-human T cell intracellular antigen 1 (TIA-1) (Beckman Coulter, Australia) (20 μg/mL), predilute rabbit anti-cow S100 (Dako, Denmark), rabbit anti-VZV IE63 polyclonal antibody (kindly provided by Prof Ravi Mahalingam, University of Colorado, Denver, CO, United States) and mouse anti-VZV gE:gI (Meridian Life Science, Saco, ME, United States). Isotype controls were mouse IgG₁, mouse IgG₂_a (Invitrogen, United States), normal rabbit and normal goat IgG (R&D systems, United States), were diluted to match primary antibody concentrations. Secondary antibodies were AlexaFluor labeled antibodies (Molecular Probes, United States) at a dilution of 1:200.

Sutherland J.P., Steain M., Buckland M.E., Rodriguez M., Cunningham A.L., Slobedman B, & Abendroth A. (2019). Persistence of a T Cell Infiltrate in Human Ganglia Years After Herpes Zoster and During Post-herpetic Neuralgia. Frontiers in Microbiology, 10, 2117.

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Multiparametric Immunofluorescence Analysis of Lymphoid Tissue

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Lymph nodes and spleens were paraffin-embedded and segmented in fragments of 2 m of thickness in a Leica microtome. Tissue sections deparaffinization, hydration and target retroviral were performed with a PT-LINK (Dako) previous to staining.
For paraffin-preserved tissue, we used rabbit anti-human CD8 (abcam), rabbit anti-human CXCR5 (GeneTex), rat anti-human CD8 (Bio-Rad), rat anti-human Granzyme B (eBioscience), mouse anti-human CD3 (Dako) and mouse anti-HIV-1 P24 (Dako), as primary antibodies; and goat antirabbit AF488 (Invitrogen), donkey anti-rat AF594 (Jackson ImmunoResearch) and donkey antimouse AF647 (ThermoFisher) as secondary antibodies. Images were taken with a Leica TCS SP5 confocal and processed with the LAS AF software. 3-D CD8 + T cell aggregations were analyzed with Imaris 9.1 software. CD8 + T cell, Granzyme B and HIV-1 P24 cell counts, co-localization and distance 2-Dimensions maps were analyzed with ImageJ software. In some cases, spleen tissue sections were also stained with hematoxylin and eosin to discriminate white (no eosin staining) and red pulp (intense eosin staining due to enrichment in erythrocytes) areas containing nucleated cells (hematoxylin stained).

Calvet‐Mirabent M., Claiborne D.T., Déruaz M., Tanno S., Serra C., Delgado‐Arévalo C., Sánchez‐Cerrillo I., Santos I.d.l., Sanz J., García‐Fraile L., Sánchez‐Madrid F., Alfranca A., Muñoz‐Fernández M.Á., Allen T.M., Buzón M.J., Balazs A.B., Vrbanac V., & Martín‐Gayo E. (2021). TAILORED DC INDUCE PROTECTIVE HIV-1 SPECIFIC POLYFUNCTIONAL CD8+ T CELLS IN THE LYMPHOID TISSUE FROM HUMANIZED BLT MICE.

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