Novus DC Differentiation and Maturation Monitoring Assay Kit, 1 Kit Item ID: NOVNBP229607

DC Differentiation and Maturation Monitoring Assay Kit Summary

Kit Type

Assay Kit

Packaging, Storage & Formulations

Storage

Store at 4C in the dark.

Applications/Dilutions

Application Notes

DC Differentiation and Maturation Monitoring Kit can be used for: Qualitative analysis of DC preparation using only a small number of cells in multicolor flow cytometry. Monitoring changes in phenotype of DCs generated by modified protocols which include change of cytokine or maturation agents. Useful tips to set-up flow staining test: 1. In DC preparations, where immature (GMCSF+IL-4) and mature DC (GMCSF+IL-4 + maturation agent) are compared, set up two separate tubes for each sample. 2. In a 3 color test, when differentiation & maturation status is analyzed, use the components of the kit with no interference of conjugate in the same channel of analysis. 3. Unstained and single color control tubes are useful for instrument set-up and compensation. 4. First time users are advised to include matched isotype controls with same conjugates as testing antibody, in staining protocols. Note: isotype control antibodies are not provided in this kit. 5. Read the application and testing protocol before starting the experiment and contact our technical team for any additional clarification.

Kit Components

Components

CD14 RPA-M1 FITC Mouse IgG1k Human 25Tests CD83 HB15e PE Mouse IgG1 k Human 25Tests CD86 IT2.2 Alexa Fluor 647 Mouse IgG2b k Human 25Tests HLA-DR L243 PE Mouse IgG2a k Human 25Tests HLA-DR L243 PerCP-Cy5.5 Mouse IgG2ak Human 25Tests

Alternate Names for DC Differentiation and Maturation Monitoring Assay Kit

• DC Phenotyping
• Dendritic cells Differentiation
• Dendritic cells Maturation
• Dendritic cells Monitoring

Background

Dendritic cells (DCs) are routinely generated from peripheral blood monocytes as precursors. The standard protocol either for clinical grade or laboratory preparations include differentiation and maturation steps with a cocktail of cytokines (GMCSF + IL-4) and maturation agents (TNFalpha, LPS and other TLR Ligands). Though this protocol generates high quality immunogenic DCs, it requires a high number of precursors to get high yields. Phenotype analysis is the first step in analyzing the quality of the DC preparation before proceeding for functional studies. Standard DC analysis includes monitoring expression of CD14, CD83, CD86 and HLA-DR antigens, as defined markers which change during differentiation and maturation. Such crucial and non-compromising analysis consumes dendritic cells, reduces sample size and consequently may limit/affect planning functional studies.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.