P slp76

Manufactured by BD

P-SLP76 is a laboratory equipment product manufactured by BD. It is a device used for the analysis and detection of specific biomolecules or cellular components. The core function of P-SLP76 is to perform analytical tasks, but a detailed description while maintaining an unbiased and factual approach is not available.

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Lab products found in correlation

Lsrii flow cytometer, by BD (1 mentions) P erk, by Cell Signaling Technology (1 mentions) P zap70, by BD (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Heat inactivated fetal calf serum, by Merck Group (1 mentions) 7 aad, by BD (1 mentions) Perk1 2, by BD (1 mentions) Cd16 biotin, by BioLegend (1 mentions) Pakt s473, by BD (1 mentions) Pplcγ2, by BD (1 mentions) Nkg2c, by Miltenyi Biotec (1 mentions)

2 protocols using p slp76

TCR Signaling Activation Dynamics

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TCR activation was performed essentially as previously reported [26 (link)]. In brief, APCs loaded with peptide at 10 μM final concentration (relevant or irrelevant) were used to activate the TCR. APCs and TCR-NK-92 were mixed at 1:1 ratio. TCR activation was stopped at different time points, 2, 5 and 15 min, followed by PFA fixation, permeabilization, barcoding and antibody staining.
Antibody staining: p-CD3zeta, p-SLP76, p-Zap70 were all from BD Biosciences, whereas p-ERK was from Cell Signaling Technologies (Leiden, The Nedherlands). Stained samples were collected on a LSRII flow cytometer (BD Biosciences). Data were analyzed using Cytobank Software (http://www.Cytobank.org), and phosphorylation levels were calculated relative to unstimulated T cells, using arcsinh transformed data.

Mensali N., Dillard P., Hebeisen M., Lorenz S., Theodossiou T., Myhre M.R., Fåne A., Gaudernack G., Kvalheim G., Myklebust J.H., Inderberg E.M, & Wälchli S. (2019). NK cells specifically TCR-dressed to kill cancer cells. EBioMedicine, 40, 106-117.

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Multiparametric NK Cell Profiling

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Antibodies were purchased for CD16 biotin from BioLegend; LFA-1 open conformation isoform was from Abcam; pZAP/Syk, pS6, pSLP76, pAKT (S473), pPLCγ2, pERK1/2, and NF-κB pp65 were from BD; KIR2DL1/S1 was from Miltenyi; KIR2DL2/3/S2 was from Beckman Coulter; KIR3DL1/2 was from BioLegend; CD57 was from BioLegend; NKG2A was from Beckman Coulter; NKG2C was from Miltenyi; CD56 was from Beckman Coulter; CD16 was from BioLegend; 7AAD was from BD; dead-cell marker was from Life Technologies; and CD107a was from BioLegend. Pacific Orange and Blue Succinimidyl Ester were bought from Thermo Fisher Scientific. 5-Methylthioadenosine (MTA) and 3-deazaadenosine (3-Deaza) were purchased from Sigma-Aldrich, 5-azacytidine (5-Aza) was from Biomol/Cayman, and 2-chloroadenosine (CADO) and EPZ015666 (EZH) were from Sigma. Avidin was purchased from Sigma. K562 cell line from ATCC was cultured in RPMI 1640 media with antibiotics (penicillin/streptomycin; Invitrogen) and 10% heat-inactivated fetal calf serum (Sigma) at 37°C.

Jacobs B., Schlögl S., Strobl C.D., Völkl S., Stoll A., Mougiakakos D., Malmberg K.J., Mackensen A, & Aigner M. (2020). The Oncometabolite 5′-Deoxy-5′-Methylthioadenosine Blocks Multiple Signaling Pathways of NK Cell Activation. Frontiers in Immunology, 11, 2128.

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