6 Capture Antibody Preparation
This procedure prepares all the capture antibodies, to be ready for coupling them to the MagPlex beads and storing for future use.
6.1 Capture Antibodies
| Manufacturer | Part No. | Pack size | PRODUCT DESCRIPTION | Component |
|---|---|---|---|---|
| bio-techne | H00054210-M04 | 100 µg | TREM1 Antibody (2E2) | Capture |
| bio-techne | MAB206-100 | 100 µg | Human/Primate IL-6 MAb (Clone 6708) | Capture |
| bio-techne | MAB2172-100 | 100 µg | Human IL-10 MAb (Clone 127107) | Capture |
| bio-techne | MAB225-100 | 100 µg | Human TNF RI/TNFRSF1A MAb (Clone 16803) | Capture |
| bio-techne | MAB25991-100 | 100 µg | Human/Primate Chitinase 3-like 1 MAb (Clone 384327) | Capture |
| bio-techne | MAB266-100 | 100 µg | Human CXCL10/IP-10/CRG-2 MAb (Clone 33036) | Capture |
| bio-techne | MAB375-100 | 100 µg | Human TRAIL/TNFSF10 MAb (Clone 75411) | Capture |
| bio-techne | MAB9231-100 | 100 µg | Human Angiopoietin-1 MAb (Clone 171733) | Capture |
| bio-techne | NB110-85467 | 100 µg | Angiopoietin-2 Antibody (MM0020-1F29) | Capture |
| bio-techne | NBP2-12045 | 100 µg | Azurocidin/CAP37/HBP Antibody (MM0099-7F31) | Capture |
| Thermo | M801 | 500 µg | Anti IL-8 (clone 3IL8-H10) | Capture |
| Thermo | MA5-24914 | 100 µg | Anti MxA | Capture |
6.2 Equipment & Consumables
| Item | Supplier | Catalogue Number |
|---|---|---|
| Pipettes and tips for 100-200 µL | Any suitable | |
| 0.2 µm syringe filter or bottle filter unit | Any suitable | |
| Syringe, 50 mL, sterile | Any suitable | |
| 50 mL tube, sterile | Any suitable | |
| Protein low-bind microtubes | Eppendorf | 0030108116 |
| Labels or permanent marker pen | Any suitable | |
| Phosphate buffered saline (PBS) | Sigma | P4417 |
6.3 Protocol
- Make up PBS as per the Buffers and Reagents protocol.
- Set out and label 10 microtubes per capture antibody.
- Remove antibodies from freezer storage.
- Add 100 µL of the PBS to each lyophilised antibody (anti IL-8 and anti MxA are already liquid at 1 mg/ml so do not need any liquid adding).
- Allow to rehydrate for a few minutes, vortex gently then spin liquid down briefly.
- Aliquot all antibodies into 10 µL volumes, or 50 µl volumes for IL-8.
- Store at -20°C or below.
All capture antibodies are now at the same stock concentration of 1 mg/mL.