Cd4 clone sk3

Manufactured by Thermo Fisher Scientific

The CD4 (clone SK3) is a lab equipment product designed for use in flow cytometry applications. It is a monoclonal antibody that specifically binds to the CD4 surface antigen expressed on certain types of T cells. The core function of this product is to facilitate the identification and enumeration of CD4+ T cells in biological samples.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

CD4 (SK3; (2 citations) PE/Cy7-conjugated CD4 (clone SK3; (2 citations)

3 protocols using cd4 clone sk3

Multiparameter Flow Cytometry of PBMCs

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

The phenotype of PBMCs cell subsets was evaluated by multiparameter flow cytometry. Cells were incubated in the dark for 20 min RT with the following panel of anti-human fluorescent labelled antibodies: CD3 (clone HIT3a, APC conjugated, BioLegend), CD4 (clone SK3, APC conjugated, eBiosciences), CD8 (clone SK1, APC conjugated, BD Biosciences), CD14 (clone 61D3, FITC conjugated, eBiosciences), CD25 (clone BC96, Alexa Fluor 488 conjugated, eBiosciences), CD69 (clone FN50, PE conjugated, eBiosciences), CD45 (clone L48, PerCyP conjugated, BD Biosciences), Foxp3 (clone PCH101, PE conjugatated, eBiosciences), IL-1β (clone CRM56, FITC conjugated, eBiosciences), IL-6 (clone MQ2-13A5, FITC conjugated, eBiosciences), IL-10 (clone JES3-9D7, Alexa Fluor 488 conjugated, eBiosciences), HLA-DR (clone L243, APC.Cy7 conjugated, BD Biosciences), CD33 (clone P67.6, PE conjugated, BD Biosciences), CD11b (clone D12, APC conjugated, BD Biosciences), CD14 (clone MɸP9, Brilliant Violet 421 conjugated, BD Biosciences) and Arginase-1 (ARG-1, FITC-conjugated, R&D Systems). Isotype controls were used for each experiment. After incubation, cells were again washed, resuspended in flow buffer and analyzed using FACSCalibur and FACSCanto II cytometers (BD, Biosciences). At least 5 × 10⁴ events were collected, and the data was analyzed using Summit software (Summit Group Software).

Domenis R., Cesselli D., Toffoletto B., Bourkoula E., Caponnetto F., Manini I., Beltrami A.P., Ius T., Skrap M., Di Loreto C, & Gri G. (2017). Systemic T Cells Immunosuppression of Glioma Stem Cell-Derived Exosomes Is Mediated by Monocytic Myeloid-Derived Suppressor Cells. PLoS ONE, 12(1), e0169932.

+ Open protocol

+ Expand

Neoantigen-reactive CD4+ T-cell Isolation

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

Cryopreserved PBMC were thawed and cultured at 2×10⁶ cells/mL in a 24-well plate with anti-CD40 at 500 ng/mL and each of the five neoantigen peptides as well as a control HIV NEF (RYPLTFGWCF) peptide at 1 µg/mL overnight at 37°C. After 16 hours, cells were stained for with LIVE/DEAD Fixable Violet Dead Cell Stain (Invitrogen), followed by lineage markers CD14 (clone M5E2, Biolegend), CD19 (clone HIB19, Biolegend), CD4 (clone SK3, eBioscience), CD8 (clone 3B5, Invitrogen) and activation-induced markers CD69 (clone L78, BD Biosciences), CD137 (clone 4-1BB, Biolegend) and CD154 (clone TRAP1, BD Bioscience). Single neoantigen-reactive CD4 cells were sorted into RNAlater (Thermo Fisher) based on coexpression of CD69, CD137 and CD154 on a BD FACSAria II (BD Biosciences).

Church C., Pulliam T., Longino N., Park S.Y., Smythe K.S., Makarov V., Riaz N., Jing L., Amezquita R., Campbell J.S., Gottardo R., Pierce R.H., Choi J., Chan T.A., Koelle D.M, & Nghiem P. (2022). Transcriptional and functional analyses of neoantigen-specific CD4 T cells during a profound response to anti-PD-L1 in metastatic Merkel cell carcinoma. Journal for Immunotherapy of Cancer, 10(9), e005328.

+ Open protocol

+ Expand

Flow Cytometry Analysis of T-Cell Markers

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

After 24 h in culture, cells were washed in PBS/0.5% FCS/0.05% NaN₃ (Sigma–Aldrich). Cells were incubated with antihuman mononuclear antibodies CD4 (clone SK3, phycoerythrin [PE]–Cyanine5.5 conjugated), CD25 (clone 2A3, PE conjugated), and CD69 (clone FN50, eFluor450 conjugated; all eBioscience) at RT for 15 min. PRBCs were lysed with BD FACS lysing solution for 10 min and washed with PBS/0.5% FCS/0.05% NaN₃. Cells were analyzed on a BD FACS Canto II flow cytometer and analyzed using the FlowJo software (version 10; BD Biosciences).

Gerner M.C., Bileck A., Janker L., Ziegler L.S., Öhlinger T., Raeven P., Müllner E.W., Salzer U., Gerner C., Schmetterer K.G, & Baron D.M. (2021). Packed red blood cells inhibit T-cell activation via ROS-dependent signaling pathways. The Journal of Biological Chemistry, 296, 100487.

+ 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!