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Cd3 alexa 488

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CD3 Alexa 488 is a fluorescently-labeled antibody that binds to the CD3 antigen, which is a component of the T cell receptor complex. It can be used for the identification and quantification of T cells in flow cytometry applications.

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Lab products found in correlation

Cd4 alexa 594, by BioLegend (3 mentions) Cd8 alexa 647, by BioLegend (3 mentions) Hoechst dye, by Merck Group (3 mentions) Cd4 pe cy7, by BioLegend (3 mentions) Fixable aqua dead cell stain, by Thermo Fisher Scientific (2 mentions) Fix lyse solution, by Thermo Fisher Scientific (2 mentions) Cd8b precp cy5, by BioLegend (2 mentions) Ack lysis buffer, by Thermo Fisher Scientific (2 mentions) Ltf 2, by BioXCell (2 mentions) Cd45 buv395, by BD (2 mentions) Cd69 percpcy5, by BioLegend (1 mentions) Ready set go elisa kit, by Thermo Fisher Scientific (1 mentions) Cd86 percp cy5, by BioLegend (1 mentions) Anti cd11c alexa488, by BioLegend (1 mentions) Cd19 alexa647, by BioLegend (1 mentions) Cd80 alexa647, by BioLegend (1 mentions) Facscalibur flow cytometry, by BD (1 mentions) Corn oil, by Merck Group (1 mentions) Ch223191, by Merck Group (1 mentions) Fixation permeabilization kit, by BD (1 mentions)

Close spelling variants of this product

Alexa 488 (13 citations) Alexa Fluor 488 anti-human CD3 (2 citations) Alexa Fluor 488 anti-mouse CD3 (2 citations)

7 protocols using cd3 alexa 488

1

Immunophenotyping of Tumor-Infiltrating T Cells

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Resected tumors were fixed for 2 h in 2% paraformaldehyde and incubated in 30% sucrose overnight. Sections were cut (5 µm) and stained with combined primary antibodies CD3 Alexa 488 (100,212, Biolegend), CD4 Alexa 594 (100,446, Biolegend), and CD8 Alexa 647 (100,727, Biolegend) and nuclei were labeled with Hoechst dye (bis benzimide, Sigma B-2283; 1 mg/100 ml in dH20). Images were acquired digitally from nine fields under each condition. Density of positive cells was evaluated by automated image analysis using Nikon Elements (Nikon Instruments Inc, Melville, NY). Percentage of CD3+ T cells, CD3+CD4+, and CD3+CD8+ T cells per area was calculated using the number of cells positive for the antibody versus the total number of cells. Student’s t-test was used to analyze statistical significance.
Yang M., Giehl E., Feng C., Feist M., Chen H., Dai E., Liu Z., Ma C., Ravindranathan R., Bartlett D.L., Lu B, & Guo Z.S. (2021). IL-36γ-armed oncolytic virus exerts superior efficacy through induction of potent adaptive antitumor immunity. Cancer Immunology, Immunotherapy, 70(9), 2467-2481.
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2

Depletion of CD8+ T Cells in ZIKV Challenge

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To deplete CD8+ T cells, anti-CD8α (BioXCell; clone YTS169.4; 500 μg) or an isotype control (BioXCell; clone LTF-2; 500 μg) was administered to immunized mice by intraperitoneal injection at days −7, −3, +1, +5 and +12. Mice then were challenged with 3 × 105 FFU of mouse-adapted ZIKV Dakar (41525) at day 0 that was preceded by intraperitoneal inoculation of anti-ifnar1 blocking mAb. To confirm immune cell depletion, peripheral blood was collected at 7 dpi followed by erythrocyte lysis with ACK lysis buffer (GIBCO) and resuspension in RPMI supplemented with 10% heat-inactivated FBS. Cells were blocked for FcγR binding and stained with CD45 BUV395 (BD BioSciences clone30-F11), CD3 Alexa488 (BioLegend clone1452C11), CD4 PE-Cy7 (BioLegend clone GK1.5), CD8b PreCP-Cy5.5 (BioLegend clone YTS156.7.7), and Fixable Aqua Dead Cell Stain (Invitrogen, L34966). Subsequently, cells were fixed by Fix/Lyse solution (eBioSciences 00–5333). Datasets were acquired on a LSRII flow cytometer and analyzed using FlowJo software X 10.0.7.
Hassan A.O., Dmitriev I.P., Kashentseva E.A., Zhao H., Brough D.E., Fremont D.H., Curiel D.T, & Diamond M.S. (2019). A Gorilla Adenovirus-Based Vaccine against Zika Virus Induces Durable Immunity and Confers Protection in Pregnancy. Cell reports, 28(10), 2634-2646.e4.
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3

Depletion of CD8+ T Cells in ZIKV Challenge

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To deplete CD8+ T cells, anti-CD8α (BioXCell; clone YTS169.4; 500 μg) or an isotype control (BioXCell; clone LTF-2; 500 μg) was administered to immunized mice by intraperitoneal injection at days −7, −3, +1, +5 and +12. Mice then were challenged with 3 × 105 FFU of mouse-adapted ZIKV Dakar (41525) at day 0 that was preceded by intraperitoneal inoculation of anti-ifnar1 blocking mAb. To confirm immune cell depletion, peripheral blood was collected at 7 dpi followed by erythrocyte lysis with ACK lysis buffer (GIBCO) and resuspension in RPMI supplemented with 10% heat-inactivated FBS. Cells were blocked for FcγR binding and stained with CD45 BUV395 (BD BioSciences clone30-F11), CD3 Alexa488 (BioLegend clone1452C11), CD4 PE-Cy7 (BioLegend clone GK1.5), CD8b PreCP-Cy5.5 (BioLegend clone YTS156.7.7), and Fixable Aqua Dead Cell Stain (Invitrogen, L34966). Subsequently, cells were fixed by Fix/Lyse solution (eBioSciences 00–5333). Datasets were acquired on a LSRII flow cytometer and analyzed using FlowJo software X 10.0.7.
Hassan A.O., Dmitriev I.P., Kashentseva E.A., Zhao H., Brough D.E., Fremont D.H., Curiel D.T, & Diamond M.S. (2019). A Gorilla Adenovirus-Based Vaccine against Zika Virus Induces Durable Immunity and Confers Protection in Pregnancy. Cell reports, 28(10), 2634-2646.e4.
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4

Immunomodulatory Effects of GD5 and DOX

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One week after the tumor injection, the tumor-bearing mice were treated intraperitoneally with 2.5 mg/kg GD5 or 3.5 mg/kg DOX. After 2 and 4 h, the spleens were collected, and the splenocytes were prepared according to the manufacturer's instructions (BioLegend). Approximately 1 × 106 cells were stained with surface antibodies and analyzed by FACSCalibur flow cytometry (BD Biosciences). The antibodies including anti-CD11c-Alexa488, -CD86-PerCP/Cy5.5, -CD80-Alexa647, -CD3-Alexa488, -CD19-Alexa647 and -CD69-PerCP/Cy5.5 were purchased from BioLegend.
Sera samples were collected at 2, 4 or 24 h from the same administrated mice and stored at -20°C. ELISA was performed using a Ready-SET-Go!® ELISA kit (eBioscience).
Gao D., Li W., Wang W., Cai Y., Wang Y., Luo X, & Wei C.C. (2017). Synergy of purine-scaffold TLR7 agonist with doxorubicin on systemic inhibition of lymphoma in mouse model. Journal of Cancer, 8(16), 3183-3189.
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5

Immunomodulatory Effects of TCDD and FICZ

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TCDD was kindly provided by Dr. Steve Safe (Institute of Biosciences & Technology, Texas A&M Health Sciences Center, College Station, Texas). FICZ was purchased from Enzo Life Sciences (Farmingdale, NY). Both TCDD and FICZ dissolved in DMSO were used for in vitro studies and dissolved in corn oil used for in vivo studies. mBSA and corn oil were purchased from Sigma-Aldrich (St. Louis, MO). RPMI 1640, L-Glutamine, penicillin-streptomycin, HEPES, PBS, and FBS were purchased from Invitrogen Life Technologies (Carlsbad, CA). AhR antagonist (CH223191) was purchased from Sigma-Aldrich, N.C. Fluorophore labeled monoclonal antibodies (mAbs) such as CD3-Alexa 488, CD4-PE/Cy7, CD8-Alexa 700, Foxp3-APC, IL-17-FITC, TGF-β1- PerCP-Cy5.5, IL10-PE, and Helois- PE–Dazzle, used for the flow cytometry, were purchased from Bio Legend (San Diego, CA) and Thermo fisher (Grand Island, NY). For intracellular staining, we used fixation/permeabilization kits from BD Biosciences for IL-17, and Foxp3 fix/perm buffer from Thermo fisher (Grand Island, NY).
Abdulla O.A., Neamah W., Sultan M., Chatterjee S., Singh N., Nagarkatti M, & Nagarkatti P. (2021). AhR Ligands Differentially Regulate miRNA-132 Which Targets HMGB1 and to Control the Differentiation of Tregs and Th-17 Cells During Delayed-Type Hypersensitivity Response. Frontiers in Immunology, 12, 635903.
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6

Quantifying Tumor-Infiltrating T-Cell Subsets

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Resected tumors were fixed for 2 h in 2% Paraformaldehyde and incubated in 30% sucrose overnight. Sections were cut (5 μm) and stained with combined primary antibodies CD3 Alexa 488 (100212, BioLegend), CD4 Alexa 594 (100446, BioLegend) and CD8 Alexa 647 (100727, BioLegend) and nuclei were labeled with Hoechst dye (bis benzimide, Sigma B-2283–1 mg/100 ml in dH20). Images were acquired digitally from 9 fields under each condition. Density of positive cells was evaluated by automated image analysis using Nikon Elements (Nikon Instruments Inc, Melville, NY). Percentage of CD3+ T cells, CD3+CD4+, CD3+CD8+ T cells per area has been calculated by number of cells positive for the antibody versus the total number of cells.
Feist M., Zhu Z., Dai E., Ma C., Liu Z., Giehl E., Ravindranathan R., Kowalsky S.J., Obermajer N., Kammula U.S., Lee A.J., Lotze M.T., Guo Z.S, & Bartlett D.L. (2020). Oncolytic Virus Promotes Tumor-Reactive Infiltrating Lymphocytes for Adoptive Cell Therapy. Cancer gene therapy, 28(1-2), 98-111.
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7

Quantifying Tumor-Infiltrating T-Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols
Resected tumors were fixed for 2 h in 2% Paraformaldehyde and incubated in 30% sucrose overnight. Sections were cut (5 μm) and stained with combined primary antibodies CD3 Alexa 488 (100212, BioLegend), CD4 Alexa 594 (100446, BioLegend) and CD8 Alexa 647 (100727, BioLegend) and nuclei were labeled with Hoechst dye (bis benzimide, Sigma B-2283–1 mg/100 ml in dH20). Images were acquired digitally from 9 fields under each condition. Density of positive cells was evaluated by automated image analysis using Nikon Elements (Nikon Instruments Inc, Melville, NY). Percentage of CD3+ T cells, CD3+CD4+, CD3+CD8+ T cells per area has been calculated by number of cells positive for the antibody versus the total number of cells.
Feist M., Zhu Z., Dai E., Ma C., Liu Z., Giehl E., Ravindranathan R., Kowalsky S.J., Obermajer N., Kammula U.S., Lee A.J., Lotze M.T., Guo Z.S, & Bartlett D.L. (2020). Oncolytic Virus Promotes Tumor-Reactive Infiltrating Lymphocytes for Adoptive Cell Therapy. Cancer gene therapy, 28(1-2), 98-111.
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