Description
The BD® OMICS-Guard Sample Preservation Buffer is a unique reagent that preserves both mRNA and protein epitopes of biological samples without the use of traditional cross-linking and harsh fixatives. The BD® OMICS-Guard Buffer is proven to preserve biological samples for both proteomic and transcriptomic studies, allowing the detection of surface and/or intracellular protein targets without compromising single-cell RNA sequencing data. The BD® OMICS-Guard Buffer can be used as a stand-alone product to preserve isolated cells, whole blood, and bulk tissues for up to 72 hours at 4°C prior to many single-cell applications (see below), or in conjunction with the BD Rhapsody™ Intracellular AbSeq Buffer Kit, which allows cells to be stored up to 24 hours at 4°C prior to permeabilization.
For stand-alone use, not in conjunction with the BD Rhapsody™ Intracellular AbSeq Buffer Kit, preserved samples can be processed and used for flow cytometry, or many multiomic single-cell workflows including whole transcriptome analysis, targeted gene analysis, or TCR/BCR full length profiling. For a current list of supported applications, contact your local Field Application Scientist (FAS). There is also an option of adding cell-surface protein profiling with these assays using BD® AbSeq Antibody-Oligo reagents. Staining with BD® AbSeq Antibody-Oligos is flexible, as cells can be stained prior to, or after preservation. If BD® AbSeq staining is done after preservation, the addition of the BD® AbSeq Enhancer is required for optimal performance.
For use with the BD Rhapsody™ Intracellular AbSeq Buffer Kit to detect mRNA and intracellular proteins, preserved cells can be safely permeabilized with Perm Buffer from the BD Rhapsody™ Intracellular AbSeq Buffer Kit without the risk of losing intracellular molecules, while concurrently allowing Intracellular BD® AbSeq Antibody-Oligos access to the interior of the cell.
Preparation And Storage
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.
Product Notices
- For U.S. patents that may apply, see bd.com/patents.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Companion Products
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.