Streptavidin-Xtra™ IF594
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Correction Factor (260 nm) | 0.34 |
Correction Factor (280 nm) | 0.15 |
Extinction coefficient (cm -1 M -1) | 1000001 |
Excitation (nm) | 568 |
Emission (nm) | 587 |
Quantum yield | 0.571 |
Intended use | Research Use Only (RUO) |
Overview | SDSProtocol |
Correction Factor (260 nm) 0.34 | Correction Factor (280 nm) 0.15 | Extinction coefficient (cm -1 M -1) 1000001 | Excitation (nm) 568 | Emission (nm) 587 | Quantum yield 0.571 |
Platform
Flow cytometer
Excitation | 561 nm laser |
Emission | 610/20 nm filter |
Instrument specification(s) | PE-Texas Red channel |
Fluorescence microscope
Excitation | Cy3/TRITC filter set |
Emission | Cy3/TRITC filter set |
Recommended plate | Black wall/clear bottom |
Example protocol
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Add 100 µL (For Cat# 46004) or 1 mL (For Cat# 46005) of ddH2O to the vial to make a 1 mg/mL stock solution.
Note: This constituted stock solution will be in PBS with 0.2% BSA.
PREPARATION OF WORKING SOLUTION
For IF, the suggested staining concentration is 1-5 ug/ml. For FACS, the suggested concentration is at 0.1-0.5 ug / 100 uL / million cells in the staining buffer.
Note: PBS + 0.1% BSA can be used as a staining buffer.
Note: For the best performance of each application, the optimal concentration of this reagent needs to be carefully determined.
Note: The suggested working dilution is provided as a guide only. It is recommended that the users titrate the product in their tests using proper positive and negative controls.
SAMPLE EXPERIMENTAL PROTOCOL
- Block and treat the samples with antibodies of interest as per the manufacturer's recommendations.
Add biotin-conjugated secondary antibody working solution in the samples at appropriate concentration and duration.
Note: Please verify the compatibility and type of your biotin conjugated antibody with the primary antibody used in the experiment. For example, If the primary antibody is a mouse antibody, then a goat Anti-Mouse antibody bound with biotin can be used for the assay.
- Incubate the cells with Streptavidin-Xtra™ working solution at room temperature for 30 minutes to 1 hour. Note: Optimal time for incubation needs to be determined carefully.
- Remove the working solution and resuspend the cells in your choice of buffer.
- Take the image using the fluorescence microscope or record the intensity using flow cytometer.
Spectrum
Spectral properties
Correction Factor (260 nm) | 0.34 |
Correction Factor (280 nm) | 0.15 |
Extinction coefficient (cm -1 M -1) | 1000001 |
Excitation (nm) | 568 |
Emission (nm) | 587 |
Quantum yield | 0.571 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
Streptavidin-Xtra™ IF488 | 491 | 516 | 750001 | 0.91 | 0.21 | 0.11 |
Streptavidin-Xtra™ IF555 | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
Streptavidin-Xtra™ IF647 | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
Images
Hela cells were fixed with 4% paraformaldehyde for 30 minutes, permeabilized with 0.02% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour. Fixed Hela cells were then stained with 1 µg/mL alpha Tubulin Mouse Monoclonal Antibody for 1 hour at room temperature, followed by GxM IgG-biotin (Cat# 16729) stain and then visualized with Streptavidin-Xtra™ iFluor 594 and Streptavidin-Alexa Fluor™ 594. Cell nuclei were stained with Hoechst 33342 (Blue, Cat#17535).
Citations
Authors: Wei, Y. P., Liu, X. P., Mao, C. J., Niu, H. L., Song, J. M., Jin, B. K.
Journal: Biosens Bioelectron (2018): 99-103
Authors: Yang, Z., Lan, Q., Li, J., Wu, J., Tang, Y., Hu, X.
Journal: Biosens Bioelectron (2017): 312-318
Authors: Hao, K., He, Y., Lu, H., Pu, S., Zhang, Y., Dong, H., Zhang, X.
Journal: Anal Chim Acta (2017): 114-120
Authors: Mishina, M., Minamihata, K., Moriyama, K., Nagamune, T.
Journal: Biomacromolecules (2017): 311
Authors: Guo, Q., Han, J. J., Shan, S., Liu, D. F., Wu, S. S., Xiong, Y. H., Lai, W. H.
Journal: Biosens Bioelectron (2016): 990-995
Authors: Chen, J., Liu, Y., Ji, X., He, Z.
Journal: Biosens Bioelectron (2016): 221-8
Authors: Lakshmipriya, T., Gopinath, S. C., Tang, T. H.
Journal: PLoS One (2016): e0151153
Authors: Mishina, M., Minamihata, K., Moriyama, K., Nagamune, T.
Journal: Biomacromolecules (2016): 1978-84
Authors: Mortberg, A., Meinke, S., Berg, P., Killie, M. K., Kjeldsen-Kragh, J., Jaras, K., Refsum, E., Hoglund, P., Wikman, A.
Journal: J Immunol Methods (2016): 9-15
Authors: Wu, D., Wang, Y., Zhang, Y., Ma, H., Pang, X., Hu, L., Du, B., Wei, Q.
Journal: Biosens Bioelectron (2016): 9-13