Description
The BD Rhapsody™ Intracellular AbSeq Buffer Kit allows users to stain for intracellular antigens using intracellular BD® AbSeq Antibody-Oligos (IC-AbSeq) for profiling with the BD Rhapsody™ System. Each intracellular BD® AbSeq Antibody is conjugated to an antibody-specific oligonucleotide barcode for profiling alongside surface protein and mRNA expression. This product provides reagents to preserve, permeabilize, and stain cells with intracellular BD® AbSeq Antibody-Oligos (sold separately). Prior to beginning the intracellular CITE-seq protocol, surface antigen staining should be performed using one of the following protocols: BD Rhapsody™ System Single-Cell Labeling with BD® AbSeq Ab-Oligos (1 plex to 40 plex) Protocol 23-24262 or BD Rhapsody™ System Single-Cell Labeling with the BD® Single-Cell Multiplexing Kit and BD® AbSeq Ab-Oligos 23-21339 to support single-cell labeling with BD® AbSeq Ab-Oligos without and with sample multiplexing, respectively. These protocols have been tested for use with up to 40 surface markers. The following protocols, BD Rhapsody™ System Single-Cell Labeling with BD® AbSeq Ab-Oligos (41 plex to 100 plex) protocol, 23-22314 or BD Rhapsody™ System Single-Cell Labeling with BD® Single-Cell Multiplexing Kit and BD® AbSeq Ab-Oligos protocol, 23-22354, are available for use with high-plex experiments, however, additional optimization may be required. See "Recommended Assay Procedure" for notes on how to use BD Rhapsody™ System Single-Cell Labeling with BD® AbSeq Ab-Oligos for Intracellular CITE-seq Protocol 23-24464 in conjunction.
After surface staining with one of the protocols listed above, users will preserve cells using BD® OMICS-Guard Sample Preservation Buffer (sold separately), permeabilize on ice with Perm Buffer (included in this kit), and stain with intracellular BD® AbSeq Antibody-Oligos (sold separately).
Immediately following this protocol users will proceed to the Single-Cell Capture and cDNA Synthesis step using one of the protocols listed in the "Recommended Assay Procedure" section.
Preparation And Storage
Store all kit components at -20 °C.
Recommended Assay Procedures
Caution: Perm Buffer contains methanol.
Base Buffer contains Bovine Serum Albumin (BSA).
Refer to the protocol BD Rhapsody™ System Single-Cell Labeling with BD® AbSeq Ab-Oligos for Intracellular CITE-seq 23-24464. Before using BD Rhapsody™ intracellular reagents, refer to the BD Rhapsody™ HT Xpress Single-Cell Analysis System Instrument User Guide 23-24257 or the BD Rhapsody™ HT Xpress System Instrument User Guide for Scanner-free workflow 23-24256. Prior to intracellular staining, prepare cells according to the following protocols:
· BD Rhapsody™ System Preparing Single-Cell Suspensions Protocol 23-24126
· BD Rhapsody™ System Single-Cell Labeling with the BD® Single-Cell Multiplexing Kit and BD® AbSeq Ab-Oligos (1 plex to 40 plex) Protocol 23-21339 or BD Rhapsody™ System Single-Cell Labeling with BD® AbSeq Ab-Oligos (1 plex to 40 plex) Protocol 23-24262 for up to 40-plex or BD Rhapsody™ System Single-Cell Labeling with BD® AbSeq Ab-Oligos (41 plex to 100 plex) protocol, 23-22314 or BD Rhapsody™ System Single-Cell Labeling with BD® Single-Cell Multiplexing Kit and BD® AbSeq Ab-Oligos protocol, 23-22354 for high-plex experiments.
Immediately after staining cells with the intracellular assay kit, one of the following protocols is required for use with the BD Rhapsody™ System:
· BD Rhapsody™ System Single-Cell Capture and cDNA Synthesis with BD Rhapsody™ Single-Cell Analysis System Protocol 23-22951
· BD Rhapsody™ System Single-Cell Capture and cDNA Synthesis with BD Rhapsody™ Express Single-Cell Analysis System Protocol 23-22952
· BD Rhapsody™ HT Single-Cell Analysis System Single-Cell Capture and cDNA Synthesis Protocol 23-24252
BD Rhapsody™ enhanced capture beads with cDNA can be stored or up to 3 months for use with the BD Rhapsody™ System mRNA Whole Transcriptome Analysis (WTA) assay. Refer to one of the following protocols:
· BD Rhapsody™ System mRNA Whole Transcriptome Analysis (WTA) and BD® AbSeq Library Preparation Protocol 23-24118
· BD Rhapsody™ System mRNA Whole Transcriptome Analysis (WTA), BD® AbSeq, and Sample Tag Library Preparation Protocol 23-24120
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@bdscomix.bd.com.
Warning: All biological specimens and materials coming into contact with 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.
Danger: Perm Buffer contains 87.68% methanol (w/w).
Hazard statements
Highly flammable liquid and vapor.
Toxic if swallowed, in contact with skin or if inhaled.
Causes damage to the central nervous system. Route of exposure: Oral.
Precautionary statements:
Wear protective clothing / eye protection.
Wear protective gloves.
Take off immediately all contaminated clothing. Wash contaminated clothing before reuse.
Wash thoroughly after handling.
Use only outdoors or in a well-ventilated area.
Keep away from heat/sparks/open flames/hot surfaces. - No smoking.
Ground and bond container and receiving equipment. Use explosion-proof [electrical/ventilating/lighting/] equipment.
Use non-sparking tools. Take action to prevent static discharges.
Do not breathe mist/vapours/spray.
Do not eat, drink or smoke when using this product.
IF ON SKIN (or hair): Remove/Take off immediately all contaminated clothing. Rinse skin with water/shower.
IF INHALED: Remove victim to fresh air and keep at rest in a position comfortable for breathing.
IF SWALLOWED: Immediately call a POISON CENTER/doctor. Rinse mouth.
If exposed: Call a poison center/doctor.
In case of fire: Use suitable extinguishing media to extinguish.
Store in well-ventilated place. Keep cool. Store locked up. Keep container tightly closed.
Danger: Proteinase K reagent contains 2.53% Proteinase K which is a respiratory hazard.
Hazard statements:
May cause allergy or asthma symptoms or breathing difficulties if inhaled
Precautionary statements:
Prevention: Avoid breathing dust/fume/gas/mist/vapors/spray
In case of inadequate ventilation wear respiratory protection
IF INHALED: If breathing is difficult, remove person to fresh air and keep comfortable for breathing
If experiencing respiratory symptoms: Call a POISON CENTER or doctor
Disposal:
Dispose of contents/container to an approved waste disposal plant
Other information:
Causes mild skin irritation.
Product Notices
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
| Description | Quantity/Size | Part Number | EntrezGene ID |
|---|---|---|---|
| Base Buffer | 1 Each (1 ea) | 51-9022685 | N/A |
| Perm Buffer | 1 Each (1 ea) | 51-9022686 | N/A |
| Nuclease Free Water | 1 Each (1 ea) | 51-9022687 | N/A |
| 0.1M DTT, Molecular Biology Grade | 1 Each (1 ea) | 51-9022688 | N/A |
| Proteinase K, Molecular Biology Grade | 1 Each (1 ea) | 51-9022689 | N/A |
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.