BD, 570911, BD® OMICS-Guard Sample Preservation Buffer

Preparation And Storage: Preparation And Storage Store all kit components at 2 to 8°C. Do not freeze. To prevent product contamination, users are recommended to make single-use aliquots in a biosafety cabinet upon opening the bottle for the first time.
Recommended Assay Procedures: Recommended Assay Procedures Contains: Bovine Serum Albumin (BSA). Use standard laboratory safety protocols. Read and understand the safety data sheets (SDSs) before handling chemicals. To obtain SDSs, go to regdocs.bd.com or contact BD Biosciences technical support at scomix@bd.com . Warning: All biological specimens and materials contacting them are considered biohazardous. Handle as if capable of transmitting infection and dispose of with proper precautions in accordance with federal, state, and local regulations. Never pipette by mouth. Wear suitable protective clothing, eyewear, and gloves. For intracellular CITE-seq assay Add the BD® OMICS-Guard Sample Preservation Buffer to pelleted cells and gently resuspend cells by pipetting. Preserve the cells at 2-8°C for 5 minutes to 24 hours for use with intracellular CITE-seq Assay using BD® AbSeq protocol. After preservation, use a swinging-bucket centrifuge, pellet cells at 800 × g for 5 minutes at 4°C. Carefully remove and discard the supernatant without disturbing the pellet. Refer to the BD Rhapsody™ System Single-Cell Labeling with BD® AbSeq Ab-Oligos for Intracellular CITE-seq Protocol 23-24464 for recommended workflow details. For stand-alone use I. Single-cell suspension Recommended usage 10^4 to 10^7 cells per 1 mL BD® OMICS-Guard Buffer Sample preservation protocol Collect single cells/dissociated single cells in suspension and spin at 400 × g for 5 minutes. 1. Discard supernatant and resuspend the cells in the BD® OMICS-Guard Buffer as recommended above. Note: Handle the buffer-containing tubes/bottle under aseptic conditions. Cells can be stored in Eppendorf Tubes® or equivalent. 2. Place cells at 4°C for up to 72 hours. 3. After storage, spin cells at 800 × g for 5 minutes and discard the supernatant to remove the BD® OMICS-Guard Buffer. No further washing is required. 4. Resuspend the cells in desired buffer for downstream applications. II. Tissue Recommended usage 30 to 50 mg of tissue per 20 mL BD® OMICS-Guard Buffer Sample preservation protocol 1. Section tissues into pieces and immediately place them into the BD® OMICS-Guard Buffer. Note: Handle the buffer-containing tubes/bottle under aseptic conditions. 2. Place cells at 4°C for up to 72 hours. 3. After storage, dissociate the tissue by the desired method for single-cell application, spin cells at 800 × g for 5 minutes and discard the supernatant to remove the BD® OMICS-Guard Buffer. No further washing is required. 4. Resuspend the cells in desired buffer for downstream applications. III. Whole blood Recommended usage 1:1 Ratio of whole blood (with EDTA) and BD® OMICS-Guard Buffer Sample preservation protocol 1. Collect whole blood using EDTA as an anticoagulant and mix with the same volume of BD® OMICS-Guard Buffer by inverting 10 times. Note: It may be possible to use other anticoagulants, though they have not been tested. 2. Place the whole blood and BD® OMICS-Guard Buffer mixture at 4°C for up to 72 hours. 3. After storage, use a magnetic red blood cell depletion kit to remove the red blood cells and isolate PBMCs and granulocytes. 4. Spin the isolated cells at 200 × g for 7 minutes to pellet nucleated cells and remove supernatant containing platelets. 5. Resuspend the leukocytes in desired buffer for downstream applications. For CITE-seq applications using BD® AbSeq Antibody-Oligos AFTER sample preservation, use of the BD® AbSeq Enhancer Kit (Cat. No. 570750) is highly recommended. BD® AbSeq Enhancers can be added to the BD Fc Block™ Reagent step or used separately prior to single-cell staining with BD® AbSeq Antibody-Oligos. Note: Staining with BD® AbSeq Antibody-Oligos BEFORE sample preservation or staining with the BD® Single-Cell Multiplexing Kit ONLY after sample preservation does not require the use of the BD® AbSeq Enhancer Kit. Staining protocol for cell suspensions (can include cells from tissues or whole blood) 1. Prepare the Human BD Fc Block™ Reagent as follows: Component Volume/sample (µL) Volume/sample with overage (µL) Stain buffer 65 78 Human BD Fc Block™ Reagent 5 6 Total 70 84 2. To the 70 µL Human BD Fc Block™ Reagent mixture, add 10 µL of each of the three BD® AbSeq Enhancers for a total of 30 µL added volume. Note: The Human BD Fc Block™ Reagent and BD® AbSeq Enhancers mixture should have a final volume of 100 µL. 3. To pelleted cells, add 100 µL final Fc Block/AbSeq Enhancer mix and resuspend the cell pellet. 4. Incubate the cells at room temperature for 10 minutes. 5. Add BD® AbSeq Antibody-Oligo cocktails per the standard AbSeq staining protocols (100 µL 2X BD® AbSeq Antibody-Oligo labeling Master Mix, incubate 30-60 minutes on ice). See BD Rhapsody™ System Single-Cell Labeling with BD® AbSeq Ab-Oligos (1 plex to 40 plex) Protocol (Doc ID: 23-24262) . 6. For BD Rhapsody™ Single-Cell Analysis System users, extend the lysis time in the single-cell capture workflow from 2 minutes to 5 minutes for optimal results for cells, tissues or whole blood that was previously preserved in BD® OMICS-Guard Buffer.