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Dry Human AIM Assay Cocktail Kit

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Regulatory Status
RUO
Other Names
Dry Reagents, Dry Cocktails
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Human_AIM-Assay-Cocktail-Kit_FC_1_062223
PHA-P-stimulated (3 day) human peripheral blood mononuclear cells were stained with the Dry Human AIM Assay Cocktail and gated on CD45+CD3+ cells. Data shown for indicated activation markers on y-axis include CD25+ and CD154+ characterized with CD8+ cells (top panels) or CD4+ cells (bottom panels).
  • Human_AIM-Assay-Cocktail-Kit_FC_1_062223
    PHA-P-stimulated (3 day) human peripheral blood mononuclear cells were stained with the Dry Human AIM Assay Cocktail and gated on CD45+CD3+ cells. Data shown for indicated activation markers on y-axis include CD25+ and CD154+ characterized with CD8+ cells (top panels) or CD4+ cells (bottom panels).
  • Human_AIM-Assay-Cocktail-Kit_FC_2_062223
    PHA-P-stimulated (3 day) human peripheral blood mononuclear cells were stained with the Dry Human AIM Assay Cocktail and gated on CD45+CD3+ cells. Data shown for indicated activation markers on y-axis include PD-L1+, OX-40+ and 4-1BB+ characterized with CD8+ cells (top panels) or CD4+ cells (bottom panels).
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320305 5 tests 198 CHF
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Description

Dry Human AIM (Activation Induced Markers) Assay Cocktail Kit is a unit dose cocktail for characterization of T cells. The Dry Human AIM Assay Cocktail Kit may be used as a stand-alone or as a backbone to which additional antibodies may be added for specific applications. The kit contains 5 tests of the Human AIM Assay Cocktail (1 test / tube) and 1 set of compensation controls with lot-matched antibodies.

The Dry Human AIM Assay Cocktail contains: 

Spark Violet™ 538 anti-human CD45 (clone HI30) 
APC/Fire™ 750 anti-human CD3 (clone UCHT1) 
Pacific Blue™ anti-human CD8 (clone SK1) 
FITC anti-human CD4 (clone A161A1) 
Spark NIR™ 685 anti-human CD25 (clone M-A251) 
PE/Dazzle™ 594 anti-human CD154 (clone 24-31) 
PerCP/Cyanine5.5 anti-human PD-L1 (clone 29E.2A3)
PE/Cyanine7 anti-human CD134 (OX-40) (clone Ber-ACT35 (ACT35))
PE anti-human CD137 (4-1BB) (clone S18014C)

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Formulation
Each kit contains the following:

5 Tubes of Dry Human AIM Assay Cocktail (1 test/tube)
1 Compensation Control Tube for Spark Violet™ 538 (1 test/tube)
1 Compensation Control Tube for APC/Fire™ 750 (1 test/tube)
1 Compensation Control Tube for Pacific Blue™ (1 test/tube)
1 Compensation Control Tube for FITC (1 test/tube)
1 Compensation Control Tube for Spark NIR™ 685 (1 test/tube)
1 Compensation Control Tube for PE/Dazzle™ 594 (1 test/tube)
1 Compensation Control Tube for PerCP/Cyanine5.5 (1 test/tube)
1 Compensation Control Tube for PE/Cyanine7 (1 test/tube)
1 Compensation Control Tube for PE (1 test/tube)

Dry cocktail is prepared from PBS containing stabilizers. Each tube can be reconstituted with 100 µL of whole blood/cell suspension or with 50 µL Cell Staining Buffer plus 50 µL of cell suspension.
Preparation
This reagent is a combination of conjugated antibodies at optimal concentrations for flow cytometry application.
Storage & Handling
The product can be stored at ambient temperature upon receipt.

Do not open the pouch until ready to use. Once opened, product can be stored at ambient temperature for up to a month in the mylar pouch.

If product is reconstituted with buffer, it can be stored in the fridge (2 - 8°C) until it is ready to use (up to 1 week).
Application

FC - Quality tested
Recommended Usage

Please refer to Application Notes

Application Notes

Dry Human AIM (Activation Induced Markers) Assay Cocktail kit is a dried down cocktail kit that aims to characterize activated T cells subsets. 

Components: 
1 pouch containing 5 tests of Dry Human AIM Assay Cocktail (1 test/tube) 
1 set of Compensation Control with lot-matched antibodies

Required Materials Not Included:
Cell Stain Buffer for Wash Steps (Cat. No. 420201 or similar)
1x RBC Lysis Buffer diluted from 10x RBC Lysis Buffer (Cat. No. 420302 or similar)
1% diluted Fixation Buffer from 4% if needed (Cat. No. 420801 or similar)

Assay Protocol:

  1. Open the Dry Human AIM Assay Cocktail Kit and the respective compensation control pouches and set up the dried down cocktail tubes to start the experiment
  2. Add 100 μL of human peripheral blood or similar cell suspension of interest into each of the dried down cocktail tubes and respective compensation controls and vortex thoroughly
  3. Incubate the samples at room temperature for 15 minutes
  4. If the cell suspension used does not need lysing, proceed to step 7. If performing Lyse wash whole blood, add 2 mL of 1x of RBC Lysis Buffer (Cat. No. 420302 or similar) to each tube and vortex thoroughly
  5. Incubate the samples at room temperature for 15 minutes
  6. Centrifuge the samples at 500 xg for 5 minutes, decant and vortex to loosen the pellet.
  7. Add 2 mL Cell Stain Buffer (Cat. No. 420201 or similar) and centrifuge the samples at 500 xg for 5 minutes, decant and vortex to loosen the pellet.
  8. Repeat step 7.
  9. Add 300 - 500 µL Cell Stain Buffer or 1% diluted Fixation Buffer (Cat. No. 420801 or similar) and acquire the samples using a flow cytometer.
RRID
AB_3097203 (BioLegend Cat. No. 320305)

Antigen Details

Distribution

Activated T Cells

Molecular Family
CD Molecules, Immune Checkpoint Receptors
Gene ID
NA
Go To Top Version: 1    Revision Date: 06.21.2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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