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- Regulatory Status
- RUO
- Other Names
- Dry Reagents, Dry Cocktails
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- Product Citations
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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). -
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).
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
- Verified Reactivity
- Human
- Formulation
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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
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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
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FC - Quality tested
- Recommended Usage
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Please refer to Application Notes
- Application Notes
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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:- 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
- 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
- Incubate the samples at room temperature for 15 minutes
- 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
- Incubate the samples at room temperature for 15 minutes
- Centrifuge the samples at 500 xg for 5 minutes, decant and vortex to loosen the pellet.
- 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.
- Repeat step 7.
- 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.
Antigen Details
- Distribution
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Activated T Cells
- Molecular Family
- CD Molecules, Immune Checkpoint Receptors
- Gene ID
- NA
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