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LEGENDScreen™

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BioLegend's LEGENDScreen™ products are lyophilized, fluorophore-conjugated antibodies provided in 96-well plates for the purpose of screening cell surface molecules on your cells of interest.

 

Why choose LEGENDScreen™?

  • Large selection of specificities per screen - Human PE Kit has 354 cell surface marker antibodies plus 10 isotype controls. Mouse PE Kit has 253 cell surface marker antibodies plus 11 isotype controls.
  • Fast and easy to follow protocol.
  • Directly conjugated antibodies at pre-titrated optimal concentrations provide reliable results.
  • Full kit with staining buffer, fixation buffer, and plate sealers.
  • Explore the Antibody Staining Data Set, an immune monitoring project that was generated using our LEGENDScreen™ Human PE Kit, and read the paper by Amir et al.

LEGENDScreen™ Mouse PE Kit (Cat. No. 700009)

 

This kit contains 253 PE-conjugated monoclonal antibodies against mouse cell surface markers, plus 11 mouse, rat, and hamster Ig isotype controls. All of the antibodies have been pre-titrated to optimum concentrations, arrayed on three 96-well plates, and lyophilized in a ready to use format. LEGENDScreen™ is the most cost-effective way to test this many directly-conjugated antibodies in a single assay.

 

This kit can be used to screen cell lines and primary cells (splenocytes, bone marrow-derived cells, and cells isolated from tissues, etc.). The workflow is simple and easy. Just reconstitute the antibodies, add your cells and then analyze. Cell staining is done directly in the plates, so no additional transfer of reconstituted antibody is required.


Blank wells on the plates allow for adding your own markers. This format allows users to screen cells of interest directly or co-stain with other markers labeled with different fluorochromes. Positive hits from the screening can be quickly identified based on the plate maps in the manual or online, with detailed clone information. Individual fluorochrome conjugated antibodies should be used to confirm the screening results. 


The LEGENDScreen™ Mouse PE Kit provides a convenient, easy to use, and powerful tool for immunology, stem cell, and cancer research.

Product Performance


The LEGENDScreen™ Mouse PE Kit was tested and compared with BioLegend's cataloged single vial liquid antibody reagents. For cell staining, cells were isolated from spleen and lymph nodes, and 3 × 105 cells were added to each well after the lyophilized antibodies were reconstituted. The cells were then stained for 20 minutes at 4°C, washed, and fixed with Fixation Buffer. The cells were then washed, resuspended in 160 µL of Cell Staining Buffer, and analyzed using an iCyt Eclipse Flow Cytometry Analyzer.

There were no significant differences in the staining patterns or median fluorescence intensity between the lyophilized product and liquid antibodies. Below are some representative data obtained when comparing the LEGENDScreen™ Mouse PE Kit vs. cataloged liquid antibodies.

 

Representative Data

 

 

Download the product manual to see more data comparisons

Tips for Successful Staining

  • Read the entire manual carefully before the experiment.
  • Plan the experiement in advance. Designate a full day for this experiment. Do not rush any step.
  • Make sure that the flow cytometer's autosampler is well maintained and working well before the experiment. If the flow cytometer does not have an autosampler, the samples from each well of the plates should be transferred to individual FACS tubes and read manually. In this case, the volume of the samples may need to be increased to avoid running dry.
  • Make sure that enough cells have been prepared for the staining. If there are not enough cells, you may choose to separate the staining into two separate experiments (e.g. stain two plates each time).
  • Depending on the application, the cell number needed for staining can be decreased. Successful staining has been done with 1 x 105 cells/well.
  • Make sure to prepare cells for machine set up and compensation. These cells should be treated the same way as the cells for staining in the plates.
  • Handle the plates with care. Keep the plates upright at all times and be careful not to knock the plates over.
  • Protect the plates from exposure to light as much as possible.
  • Use extra caution when opening the plate cover (see Plate Preparation for instructions). After the cover is removed, do not apply any sealers on the plates until the cakes are reconstituted. Applying plate sealers or any cover onto the plate before the cakes are reconstituted can result in cakes being dislodged and stuck to the cover or escaping from the wells.
  • Some cell surface markers are sensitive to enzymatic digestion. If adherent cells are being used for staining, a mild enzyme or non-enzymatic dissociation buffer should be used when possible.
  • Make sure cells are in a single cell suspension. DNase treatment is recommended to avoid clumps caused by dead cells.
  • Analyze only 70 µL of the 160 µL samples so that a second run can be performed if necessary.

 

FAQs


Is there any data analysis tool or algorithm applicable to this product?​
Currently, we do not offer any automated analysis solutions for LEGENDScreen™ kits. However, a commercial solution is available from Astrolabe Diagnostics. The Antibody Staining Data Set is an example application of the LEGENDScreen™ Human PE Kit (see Amir et al.).

What is the level of variability from one experiment to the other?
If the protocol is followed closely, the variability should be minimal. The variability should be similar to single vial antibody staining. 

How should the kit be stored?
The kit should be stored at 2 - 8°C upon receipt. Once opened, the plates must be reconstituted immediately. Reconstituted plates can be used or stored at 2 - 8°C sealed in the dark and used within a month. 

How do I request a custom LEGENDScreen™ product with only my specificities of interest?
For more info, visit: biolegend.com/custom_solutions. 

What are the guarantees regarding the lyophilized plate compared to the reconstituted plate?
Lyophilized product has a guaranteed shelf life of 6 months unopened. Reconstituted plates can be used or stored at 2 - 8°C sealed in the dark and used within a month. Be sure to properly seal the plates to prevent evaporation and shield the antibodies from light. 

I have added my own antibody solution to the lyophilized product, will the lyophilized antibody work?
Yes, as long as the fluorophores on these antibodies are compatible and proper compensation has been applied during acquisition and analysis. 

I am not going to use all the reconstituted antibody solution. Can I keep the leftover for later or re-dry the solution in the dark?
The antibody is in a one test per well format. There will not be any antibody left if the full test is used. Customers may decide to use less than the recommended volume per test, but this is not recommended and the performance is not guaranteed. Customers may also selectively transfer certain antibodies from the original plate to a new plate and use after reconstitution. If any antibody is not used after reconstitution, the plate can be sealed and store at 2 - 8°C for a month in the dark. Once reconstituted, re-drying is not recommended as this may result in a loss of signal.  

If I don't have enough cells and use less than 4 x 106/ml (3 x 105 cells/well), will it still work? 
This may work with lower numbers of total cells, but we recommend trying to keep higher concentrations of cells for faster analysis. Of course, how many cells are needed depends on the specific application. Successful staining has been done with 1 x 105 cells/well. 

Are these plates made under sterile conditions?
The plates are not sterile. You would handle them as you would handle most typical flow cytometry staining protocols. 

Can I use half or less of the plate and keep the rest for later?
Yes. Customers can use half of the plate or whatever specificities they are interested in. However, the whole plate should be reconstituted. The half plate of antibodies must be transferred to another empty plate for the staining. The remaining half must be sealed and stored at 2 - 8°C in the dark and used within a month. 

For more answers, 

LEGENDScreen™ Human PE Kit (Cat. No. 700011)


This kit contains 354 PE-conjugated monoclonal antibodies against human cell surface markers, plus 10 mouse, rat, and hamster Ig isotype controls. All of the antibodies have been pre-titrated to optimum concentrations, arrayed on four 96-well plates, and lyophilized in a ready to use format. LEGENDScreen™ is the most cost-effective way to test this many directly-conjugated antibodies in a single assay.

 

This kit can be used to screen cell lines and primary cells (PBMCs, bone marrow derived cells, and cells isolated from tissues). The workflow is simple and easy. Just reconstitute the antibodies, add your cells and then analyze. Cell staining is done directly in the plates, so no additional transfer of reconstituted antibody is required.

Blank wells on the plates allow for adding your own markers. This format allows users to screen cells of interest directly or co-stain with other markers labeled with different fluorochromes. Positive hits from the screening can be quickly identified based on the plate maps in the manual or online, with detailed clone information. Individual fluorochrome conjugated antibodies should be used to confirm the screening results. 

The LEGENDScreen™ Human PE Kit provides a convenient, easy to use, and powerful tool for immunology, stem cell, and cancer research.

Product Performance


The LEGENDScreen™ Human PE Kit was tested and compared with BioLegend's cataloged single vial liquid antibody reagents. For cell staining, human PBMCs were isolated and 3 × 105 cells were added to each well after the lyophilized antibodies were reconstituted. The cells were then stained for 20 minutes at 2 - 8°C, washed, and fixed with Fixation Buffer. The cells were then washed, resuspended in 160 µL of Cell Staining Buffer, and analyzed using a Beckton Dickenson FACS Canto II Flow Cytometry Analyzer.

The staining patterns and median fluorescence intensity between the lyophilized product and liquid antibodies are similar. Below are some representative data obtained when comparing the LEGENDScreen™ Human (PE) Kit vs. equivalent liquid antibodies.

 

Representative Data  

 

Tips for Successful Staining

 

  • Read the entire manual carefully before the experiment.
  • Plan the experiment in advance. Designate a full day for this experiment. Do not rush any step.
  • Make sure that the flow cytometer’s autosampler is well maintained and working well before the experiment. If the flow cytometer does not have an autosampler, the samples from each well of the plates should be transferred to individual FACS tubes and tube adjust the sample volume to approximately 300µl or more to avoid running dry. Alternatively, a Cluster Tube System (such as Corning catalog # 4410) and acquired manually. The cluster tube can then be transferred to a regular FACS tube for acquisition.
  • Make sure that enough cells have been prepared for the staining. If there are not enough cells, you may choose to divide the staining into two separate experiments.
  • Depending on the application, the cell number needed for staining can be decreased. Successful staining has been done with1 x 105 cells/well.
  • Make sure to prepare cells for machine setup and compensation. These cells should be treated the same way as the cells for staining in the plates.
  • Handle the plates with care. Keep the plates upright at all times and be careful not to knock the plates over.
  • Protect the plates from exposure to light as much as possible.
  • Use extra caution when opening the plate cover (see Plate Preparation for instructions). After the cover is removed, do not move or apply any sealers on the plates until the cakes are reconstituted. Applying plate sealers or any cover onto the plate before the cakes are reconstituted can result in cakes being dislodged and stuck to the cover or escaping from the wells.
  • Some cell surface markers are sensitive to enzymatic digestion. If adherent cells are being used for staining, a mild enzyme or non-enzymatic dissociation buffer should be used when possible.
  • Make sure cells are in a single cell suspension. DNase treatment is recommended to avoid clumps caused by dead cells.
  • Acquire only 70 µL of the 160 µL total volume so that a second run can be performed if necessary.

 

FAQs


Is there any data analysis tool or algorithm applicable to this product?​
Currently, we do not offer any automated analysis solutions for LEGENDScreen™ kits. However, a commercial solution is available from Astrolabe Diagnostics. The Antibody Staining Data Set is an example application of the LEGENDScreen™ Human PE Kit (see Amir et al.).

What is the level of variability from one experiment to the other?
If the protocol is followed closely, the variability should be minimal. The variability should be similar to single vial antibody staining.

How should the kit be stored?
The kit should be stored at 2 - 8°C upon receipt. Once opened, the plates must be reconstituted immediately. Reconstituted plates can be used or stored at 2 - 8°C sealed in the dark and used within a month.

How do I request a custom LEGENDScreen™ product with only my specificities of interest?
For more info, visit: biolegend.com/custom_solutions.

What are the guarantees regarding the lyophilized plate compared to the reconstituted plate?
Lyophilized product has a guaranteed shelf life of 6 months unopened. Reconstituted plates can be used or stored at 2 - 8°C sealed in the dark and used within a month. Be sure to properly seal the plates to prevent evaporation and shield the antibodies from light.

I have added my own antibody solution to the lyophilized product, will the lyophilized antibody work?
Yes, as long as the fluorophores on these antibodies are compatible and proper compensation has been applied during acquisition and analysis.

I am not going to use all the reconstituted antibody solution. Can I keep the leftover for later or re-dry the solution in the dark?
The antibody is in a one test per well format. There will not be any antibody left if the full test is used. Customers may decide to use less than the recommended volume per test, but this is not recommended and the performance is not guaranteed. Customers may also selectively transfer certain antibodies from the original plate to a new plate and use after reconstitution. If any antibody is not used after reconstitution, the plate can be sealed and store at 2 - 8°C for a month in the dark. Once reconstituted, re-drying is not recommended as this may result in a loss of signal.  

If I don't have enough cells and use less than 4 x 106/ml (3 x 105 cells/well), will it still work?
This may work with lower numbers of total cells, but we recommend trying to keep higher concentrations of cells for faster analysis. Of course, how many cells are needed depends on the specific application. Successful staining has been done with 1 x 105 cells/well.

Are these plates made under sterile conditions?
The plates are not sterile. You would handle them as you would handle most typical flow cytometry staining protocols.

Can I use half or less of the plate and keep the rest for later?
Yes. Customers can use half of the plate or whatever specificities they are interested in. However, the whole plate should be reconstituted. The half plate of antibodies must be transferred to another empty plate for the staining. The remaining half must be sealed and stored at 2 - 8°C in the dark and used within a month.

For more answers,

 

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