αccr7 pecy7

Manufactured by BD

Sourced in United States

αCCR7‐PECy7 is a fluorescently labeled antibody that binds to the CCR7 receptor, which is expressed on certain immune cells. It can be used for the identification and analysis of CCR7-positive cells in flow cytometry applications.

Automatically generated - may contain errors

Lab products found in correlation

3 protocols using αccr7 pecy7

Single-Cell TCR Sequencing of Tetramer-Positive CD8+ T Cells

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

Tetramer magnetic enrichment and single‐cell PCR were undertaken as previously described.³⁷, ⁴⁷ Briefly, PBMCs from HLA‐A*03:01⁺ individuals were FcR blocked (Miltenyi Biotech) in MACS buffer (phosphate‐buffered saline (PBS)), 0.5% bovine serum albumin (BSA; Sigma‐Aldrich); and 0.2 mm EDTA (Sigma‐Aldrich) for 15 min at 4°C. PBMCs were then stained with PE‐conjugated tetramer in MACS buffer for 1 h at room temperature, washed and labelled with anti‐PE microbeads (Miltenyi Biotech) at 4°C for 30 min. Epitope‐specific cells were enriched by passing twice over a LS magnetic column (Miltenyi Biotech) and surface stained with αCD3‐BV480 (BD Biosciences), αCD8‐PerCPCy5.5 (BD Biosciences), Live/Dead‐NIR (Life Technologies), αCD14‐APCH7 (BD Biosciences), αCD4‐APCH7 (BD Biosciences), αCD19‐APCH7 (BD Biosciences), αCD27‐BV711 (BD Biosciences), αCD45RA‐FITC (BD Biosciences), αCCR7‐PECy7 (BD Biosciences) and αCD95‐BV421 (BD Biosciences) at 4°C in the dark. Cells were resuspended in sort buffer (PBS, 0.1% BSA; Gibco, CA, USA), and tetramer^high/CD8⁺ T cells were single‐cell sorted directly into Twin‐Tech PCR plates (Eppendorf, Hamburg, Germany) on an Aria Fusion (BD Biosciences) and were stored at −80°C until used. The gating strategy is shown in Supplementary figure 1c.

Nguyen A.T., Lau H.M., Sloane H., Jayasinghe D., Mifsud N.A., Chatzileontiadou D.S., Grant E.J., Szeto C, & Gras S. (2022). Homologous peptides derived from influenza A, B and C viruses induce variable CD8 + T cell responses with cross‐reactive potential. Clinical & Translational Immunology, 11(10), e1422.

+ Open protocol

+ Expand

Multiparametric Analysis of Antigen-Specific CD8+ T Cells

Cited 1 time

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

Standard and CD8-null ELA/HLA-A2 tetramers were produced as described previously (16 (link), 17 (link)). The CD8-null ELA/HLA-A2 complex incorporates a compound D227K/T228A mutation in the α3 domain that abrogates CD8 binding without impacting on the TCR docking platform (18 (link)). Directly conjugated monoclonal antibodies (mAbs) were purchased from commercial sources as follows: (i) α-CD8-APC-Cy7, α-CD45RA-V450, α-CCR7-PE-Cy7, α-CD107a-PE-Cy5, α-IFNγ-AlexaFluor700, α-TNF-PE-Cy7 and α-granzyme-B-V450 (BD Biosciences); (ii) α-CD3-ECD (Beckman Coulter); (iii) α-CD28-AlexaFluor700 (BioLegend); (iv) α-T-bet-AlexaFluor647 (eBiosciences); (v) α-MIP-1β-FITC (R&D Systems); (vi) α-IL-2-APC (Miltenyi Biotec); and (vii) α-perforin-BD48-FITC (Abcam). The amine-reactive viability dye Aqua (Life Technologies) was used to eliminate dead cells from the analysis. Intracellular staining for T-bet was performed using the Transcription Factor Buffer Set (BD Pharmingen) according to the manufacturer’s instructions. Intracellular staining for granzyme B and perforin-BD48 was compatible with this procedure. Staining with all other reagents was conducted according to standard protocols (19 (link), 20 (link)). Data were acquired using an LSR Fortessa flow cytometer (BD Biosciences) and analyzed with FlowJo software version 9.3.7 (TreeStar Inc.).

Lissina A., Briceño O., Afonso G., Larsen M., Gostick E., Price D.A., Mallone R, & Appay V. (2015). Priming of qualitatively superior human effector CD8+ T cells using TLR8 ligand combined with FLT3L. Journal of immunology (Baltimore, Md. : 1950), 196(1), 256-263.

+ Open protocol

+ Expand

Multiparameter Flow Cytometric Analysis

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

The following mAb conjugates were used at pre-titrated concentrations: (i) αCD3-APC-H7 (BD Biosciences, San Jose, CA, USA); (ii) αCD8-QD705, αCD14-Pacific Blue and αCD19-Pacific Blue (Life Technologies); (iii) αCD8-PE, αCD57-FITC and αCCR7-PE-Cy7 (BD Pharmingen, San Jose, CA, USA); and (iv) αCD27-PE-Cy5, αCD45RA-ECD and αCD45RO-ECD (Beckman Coulter, High Wycombe, UK). LIVE/DEAD Fixable Aqua and Violet Dead Cell Stain Kits (Life Technologies) were used to eliminate nonviable cells from the analysis. Peptide-major histocompatibility complex dextramers conjugated separately to APC and PE (Immudex, Copenhagen, Denmark) were used for magnetic enrichment.

Neller M.A., Ladell K., McLaren J.E., Matthews K.K., Gostick E., Pentier J.M., Dolton G., Schauenburg A.J., Koning D., Fontaine Costa A.I., Watkins T.S., Venturi V., Smith C., Khanna R., Miners K., Clement M., Wooldridge L., Cole D.K., van Baarle D., Sewell A.K., Burrows S.R., Price D.A, & Miles J.J. (2015). Naive CD8+ T-cell precursors display structured TCR repertoires and composite antigen-driven selection dynamics. Immunology and Cell Biology, 93(7), 625-633.

+ 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!