Rhodamine B hydrazide |
カタログ番号GC44832 |
ロダミンBハイドラジドは、励起/発光スペクトルがそれぞれ510 / 578 nmの水溶性蛍光プローブです。
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 74317-53-6
Sample solution is provided at 25 µL, 10mM.
評判の良い論文で引用されたGlpBio製品
Product Documents
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Protocol
This plan only provides a guide, please modify it to meet your specific needs.
1. Prepare staining solution
(1) Prepare stock solution: Dissolve Rhodamine B hydrazide in DMSO and prepare a stock solution with a concentration of 5-10mM.
Note: Unused stock solution should be aliquoted and stored in the dark at -20℃ or -80℃ to avoid repeated freezing and thawing.
(2) Prepare working solution: Dilute the stock solution with a suitable buffer (such as serum-free medium or PBS) and prepare a working solution with a concentration of 5-10μM.
Note: Please adjust the concentration of the working fluid according to the actual situation and prepare it now.
2. Cell suspension staining
(1) Suspension cells: Centrifuge at 1000g for 3-5 minutes at 4℃, discard the supernatant, and wash twice with PBS for 5 minutes each time.
(2) Adherent cells: Wash twice with PBS, add trypsin to digest the cells, and centrifuge at 1000g for 3-5 minutes after digestion is completed.
(3) Add Rhodamine B hydrazide working solution to resuspend the cells, and incubate at 37℃ in the dark for 2 hours. The optimal incubation time for different cells is different, please explore by yourself according to the specific experimental needs.
(4) After the incubation, centrifuge at 1000g for 5 minutes, remove the supernatant, add PBS and wash 2-3 times, 5 minutes each time.
(5) Resuspend cells in pre-warmed serum-free cell culture medium or PBS and observe by fluorescence microscope or flow cytometry.
3. Cell adhesion staining
(1) Culture adherent cells on sterile coverslips.
(2) Remove the coverslip from the culture medium, suck out the excess culture medium, and place the coverslip in a humid environment.
(3) Add 100μL of dye working solution from one corner of the coverslip and shake gently to evenly cover all cells with the dye.
(4) Incubate at 37℃ in the dark for 2 hours. The optimal incubation time for different cells is different, please explore by yourself according to the specific experimental needs.
(5) After the incubation, discard the dye working solution and use pre-warmed culture solution to wash the coverslip 2 to 3 times.
4. Detect intracellular nitric oxide production
(1) Incubate cells with 5μM Rhodamine B hydrazide (2.5%, v/v DMSO) for 4 hours at 37℃ in the dark;
(2) Discard the dye solution and wash twice with PBS;
(3) Incubate the cells with DMEM containing 375μM SNAP (S-nitroso-N-acetyl-D,L-penicillamine, a NO releasing agent) for 2 hours at 37℃ in the dark;
(4) Wash twice with PBS, then soak the sample in PBS.
5. Microscope detection: The maximum excitation wavelength and emission wavelength of Rhodamine B hydrazide are 546/610nm respectively.
Precautions:
1) Fluorescent dyes all have quenching problems. Please avoid light as much as possible to slow down fluorescence quenching.
2) For your safety and health, please wear a lab coat and disposable gloves.
References:
[1]. Chi-Ming Wu,et,al. Sensitivity evaluation of rhodamine B hydrazide towards nitric oxide and its application for macrophage cells imaging. 2011 Dec 5;708(1-2):141-8. doi: 10.1016/j.aca.2011.10.005. Epub 2011 Oct 8.
Rhodamine B hydrazide is a water-soluble fluorescent probe with excitation/emission spectra of 510/578 nm, respectively. It has been used for the detection of copper, peroxynitrite, nitric oxide, hydrogen peroxide, glucose, diacetyl, and hemoglobin.
Cas No. | 74317-53-6 | SDF | |
Canonical SMILES | CCN(CC)C1=CC2=C(C=C1)C3(C(C=CC=C4)=C4C(N3N)=O)C5=CC=C(N(CC)CC)C=C5O2 | ||
Formula | C28H32N4O2 | M.Wt | 456.6 |
溶解度 | DMF: 30 mg/ml,DMF:PBS(pH7.2) (1:2): 0.33 mg/ml,DMSO: 20 mg/ml,Ethanol: 0.1 mg/ml | Storage | Store at -20°C |
General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. |
Complete Stock Solution Preparation Table
Prepare stock solution | |||
1 mg | 5 mg | 10 mg | |
1 mM | 2.1901 mL | 10.9505 mL | 21.901 mL |
5 mM | 0.438 mL | 2.1901 mL | 4.3802 mL |
10 mM | 0.219 mL | 1.0951 mL | 2.1901 mL |
In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Related Products
- GC12410 Bupivacaine HCl
- GC37735 Tankyrase-IN-2
- GC12536 OC000459
- GC19321 Savolitinib
- GC15612 Luotonin A
- GC36115 Ganoderenic acid E
- GC16523 Dipraglurant
- GC39738 AQC
- GC30600 4-Ethylphenol
- GC36337 Isopropylidenylacetyl-marmesin
- GC40082 Drimentine B
- GC37840 Tubulysin F
- GC44339 N-Cyclohexanecarbonylpentadecylamine
- GC14067 Prasugrel hydrochloride
- GC32859 Phorbol (4β-Phorbol)
- GC36925 Piperaquine tetraphosphate tetrahydrate
- GC38133 Tirzepatide (TFA)
レビュー
Average Rating: 5 ★★★★★ (Based on Reviews and 22 reference(s) in Google Scholar.)
GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
Required fields are marked with *