The largest database of trusted experimental protocols

Quicklysis

Manufactured by Cytognos

Quicklysis is a rapid cell lysis solution designed to prepare samples for flow cytometry analysis. It efficiently disrupts cell membranes, releasing cellular contents for subsequent staining and detection.

Automatically generated - may contain errors

4 protocols using quicklysis

1

Immune Cell Phenotyping by Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immune cells carry out some of their functions by means of cell-to-cell contact and via membrane receptors or antigens that are expressed at the cellular surface. Some molecules appear at various stages of cell differentiation or activation. In the present study, we recorded the following molecules: CD4, CD8, CD19, CD56, CD71, CD97, CD134, and CD173.
Phenotypic analysis of the various cell populations was carried out by immunofluorescence by means of flow cytometry with double or triple labeling. This technique constitutes a useful tool for cell identification and characterization. Following intravenous peripheral blood collection, 50 μl of blood were incubated for 10 minutes with 3 μl of the following combinations of monoclonal antibodies (Pharmigen):

CD4-PE + CD8-FITC + CD19-Cy5

CD8-FITC + CD56-PE

CD71-FITC + CD97-PE

CD134-FITC + CDw137-PE

Afterwards, 1 mL of a hematolysis solution (Quicklysis, Cytognos) was added and the blood solution was kept in the dark for 5 minutes, after which time 10,000 cells were subjected to cytometric analysis.
Immunofluorescence analysis was carried out by means of a Vantage (Becton Dickinson) FACS flow cytometer, equipped with an argon laser (488 nm) that excites the FITC, PE, and Cy5 fluorchromes, emitting at 520, 575, and 667 nm, respectively.
+ Open protocol
+ Expand
2

Immunophenotyping of Murine Mesenchymal Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
To confirm the phenotype of mMSC cells were stained with specific antibodies during 30 min at 4 degrees. MSC were positive for CD29, CD44, and Sca-1 and negative for CD45, CD11b, and CD14 (data not shown).
To analyze immune infiltrated cells, tumor mass was carefully washed with HBSS 1X and mechanically processed prior enzymatic digestion with 1 mg/mL of collagenase D (Roche; Catalog #11088858001). Tumor cell suspension was counted by trypan blue and 5 x 105 alive cells were labelled with specific antibodies during 30 min at 4 degrees. Red blood cells were lysed using QuickLysis (Cytognos; Catalog #CYT-QL-1) for 20 min in darkness. Used antibodies are indicated in supplementary information (Supplementary Table 1). Samples were acquired in a FacsCanto II (BD, San Jose, CA) cytometer and analyzed using FacsDiva software.
+ Open protocol
+ Expand
3

Multifaceted T Cell Phenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were labeled with fluorescent antibodies (shown in Table 1) against human to study the phenotype of different subsets of T cells (shown in the Table 2), the early activation of T cells (CD25+ CD134+) or the exhaustion of T lymphocytes by measuring the levels of inhibitory receptors.
Cell phenotype was studied in fresh samples and after 14 days of culture using the gating strategy for identifying T cell subsets shown in Supplementary Figure S1. In the case of CAR-T cell assays, the phenotype was analyzed 7 days after transduction. Staining protocol was performed at 4°C during 30 min and erythrocytes were lysed with quicklysis (Cytognos). Events were acquired using a BD FacsCanto II cytometer, and data were analyzed with FacsDiva software (BD) and FlowJo software version 10.6.1.
+ Open protocol
+ Expand
4

Tumor Cell Dissociation and Flow Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tumor cells were disaggregated into individual cells with Accumax (5 min, RT) and erythrocytes were lysed with Quicklysis (15 min, RT; Cytognos) before staining. Cells were stained with anti-CD44-FITC (ImmunoTools, Supplementary Table S1) diluted in PBS+0.5% BSA+2 mM EDTA (staining buffer) for 20 min on ice and treated with PI (5 μg/ml, Sigma-Aldrich) for 5 min on ice. After staining, cells were washed with staining buffer and analyzed by flow cytometry (FACSCalibur, Becton Dickinson) using the FlowJo software.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!