mCherry Nanobody Bead
Anti-mCherry Nanobody Immunomagnetic Beads
Synonyms
mCherry Nanobody; N/A; Anti-mCherry Nanobody Immunomagnetic Beads; mCherry Nanobody bead
Host
Alpaca
Isotype
VHH
Antibody properties
Anti-mCherry Nanobody.
Magnetic beads properties
Agarose coated superparamagnetic beads with an average particle size of 3 um
Binding capacity
0.5mL nano magnetic beads are covalently coupled with 2mg Anti-mCherry nanobody. 0.5mL Anti-mCherry immunomagnetic beads can precipitate at least 0.6mg mCherry fusion protein.
Components
0.25mL Anti-mCherry immunomagnetic beads stored in 0.75mL PBS containing preservatives.
Matters Needing Attention
1. This product is only used for scientific research by professionals, and shall not be used for clinical identification or treatment.
2. For your safety and health, please wear lab clothes and disposable gloves.
3. This product provides affinity magnetic beads in the form of suspension. The content of magnetic beads in the suspension is 25%.Before use, gently re-suspend the magnetic bead suspension, and then use it as required.
4. Do not centrifuge, freeze or dry the magnetic beads do not use sonication for the magnetic beads, and do not allow the acid treatment time of the magnetic beads to exceed 10min
5. When mixing the magnetic beads, please use the pipette to gently blow, use soft vortex, turn upside down, shaking table mixing and other methods.
6. Related reagents for supporting use shall be prepared by the laboratory itself.
2. For your safety and health, please wear lab clothes and disposable gloves.
3. This product provides affinity magnetic beads in the form of suspension. The content of magnetic beads in the suspension is 25%.Before use, gently re-suspend the magnetic bead suspension, and then use it as required.
4. Do not centrifuge, freeze or dry the magnetic beads do not use sonication for the magnetic beads, and do not allow the acid treatment time of the magnetic beads to exceed 10min
5. When mixing the magnetic beads, please use the pipette to gently blow, use soft vortex, turn upside down, shaking table mixing and other methods.
6. Related reagents for supporting use shall be prepared by the laboratory itself.
Method of Application
1. Preparation of cell lysate
1) Collecting cells
Blow the suspended cells and semi-adherent cells off the cell culture flask and transfer them into a centrifuge tube, centrifuge at 1000rpm for 5min, and discard the supernatant. Gently scrape the adherent cells off the bottle wall with a cell scraper, transfer them into a centrifuge tube together with the culture medium, centrifuge at 1000rpm for 5min, and discard the supernatant.
2) Re-suspend the cells with 1x PBS pre-cooled to 4 degree C, centrifuge at 1000rpm for 3min, and discard the supernatant. Repeat.
3) Add the corresponding volume of cell lysate according to the amount of cells, and place it on the ice for 10-20 min after repeated blowing.
Note: Generally, 1mL of cell lysis buffer can process about 0.5-1 x107 cells. To avoid degradation of that target protein, you may add protease inhibitor.
4) Treat cell lysate with ultrasonic crusher until cell lysate is clear and no longer viscous. After 30 min on ice, centrifuge at 12000 rpm for 10 min at 4 degree C. Take out the supernatant and freeze at -80 degree C.
5) If the target protein is secreted and expressed, the above treatment is not required, the supernatant of the medium can be directly collected and the following steps can be performed after concentration.
2. Detection of mCherry Tagged Protein by Immuno (co) precipitation Method
1) Gently re-suspend the Anti-mCherry magnetic beads, take 40 uL magnetic beads suspension (including 10 uL magnetic beads) put it in the centrifuge tube, add 500 uL 1x PBS, fully suspended the suspension and place on the magnetic rack for magnetic separation. After the magnetic rack is left standing for 10 seconds, discard the supernatant. Repeat this washing step twice. Note: For multiple samples, the magnetic beads can be re-suspended and then divided into several reaction tubes for reaction.
2) Add 50-200 uL eukaryotic cell lysate containing target protein, gently re-suspend magnetic beads, and incubate in a shaking table at room temperature for 2h or at 4 degree C overnight.
3) After standing on the magnetic rack for 10 seconds, transfer the supernatant to a new centrifuge tube for later use (the supernatant can be used to detect whether mCherry-tag protein remains). Add 500 uL 1xPBS, mix gently, clean the magnetic beads, perform magnetic separation, and discard the supernatant. Repeat twice.
4) Add 20 uL 1xPBS and 5 uL 5x loading buffer, boil the sample for 5min, cool it to room temperature and centrifuge.
5) Take the supernatant for SDS-PAGE test and for subsequent Western Blotting detection.
1) Collecting cells
Blow the suspended cells and semi-adherent cells off the cell culture flask and transfer them into a centrifuge tube, centrifuge at 1000rpm for 5min, and discard the supernatant. Gently scrape the adherent cells off the bottle wall with a cell scraper, transfer them into a centrifuge tube together with the culture medium, centrifuge at 1000rpm for 5min, and discard the supernatant.
2) Re-suspend the cells with 1x PBS pre-cooled to 4 degree C, centrifuge at 1000rpm for 3min, and discard the supernatant. Repeat.
3) Add the corresponding volume of cell lysate according to the amount of cells, and place it on the ice for 10-20 min after repeated blowing.
Note: Generally, 1mL of cell lysis buffer can process about 0.5-1 x107 cells. To avoid degradation of that target protein, you may add protease inhibitor.
4) Treat cell lysate with ultrasonic crusher until cell lysate is clear and no longer viscous. After 30 min on ice, centrifuge at 12000 rpm for 10 min at 4 degree C. Take out the supernatant and freeze at -80 degree C.
5) If the target protein is secreted and expressed, the above treatment is not required, the supernatant of the medium can be directly collected and the following steps can be performed after concentration.
2. Detection of mCherry Tagged Protein by Immuno (co) precipitation Method
1) Gently re-suspend the Anti-mCherry magnetic beads, take 40 uL magnetic beads suspension (including 10 uL magnetic beads) put it in the centrifuge tube, add 500 uL 1x PBS, fully suspended the suspension and place on the magnetic rack for magnetic separation. After the magnetic rack is left standing for 10 seconds, discard the supernatant. Repeat this washing step twice. Note: For multiple samples, the magnetic beads can be re-suspended and then divided into several reaction tubes for reaction.
2) Add 50-200 uL eukaryotic cell lysate containing target protein, gently re-suspend magnetic beads, and incubate in a shaking table at room temperature for 2h or at 4 degree C overnight.
3) After standing on the magnetic rack for 10 seconds, transfer the supernatant to a new centrifuge tube for later use (the supernatant can be used to detect whether mCherry-tag protein remains). Add 500 uL 1xPBS, mix gently, clean the magnetic beads, perform magnetic separation, and discard the supernatant. Repeat twice.
4) Add 20 uL 1xPBS and 5 uL 5x loading buffer, boil the sample for 5min, cool it to room temperature and centrifuge.
5) Take the supernatant for SDS-PAGE test and for subsequent Western Blotting detection.
Note
Do not centrifuge and use after mixing gently
Preparation and Storage
Store at 4 degree C for 12 months
Related Product Information for mCherry Nanobody bead
Background: mCherry, a red fluorescent protein derived from sea anemone (Discosoma sp.), is composed of 266 amino acids. Because of its color and light stability of monomer molecules, it performs better than other fluorescent protein tags. mCherry is often used as eukaryotic protein recombinant expression marker. Anti mCherry nanobody immu nomagnetic beads are made by covalently coupling high quality mCherry nanobody with magnetic beads. It has the characteristics of high binding capacity of protein, high specificity and stability, fast and convenient operation, r epeatability and can be used for immuno (co) precipitation of mCherry tagged fusion proteins.
Product Categories/Family for mCherry Nanobody bead
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Product Notes
The mCherry Nanobody (Catalog #AAA178633) is a Bead produced from Alpaca and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "mCherry Nanobody, Bead" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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