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1mg |
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5mg |
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ln Vitro |
AF488 NHS ester IgG fluorescent labeling and anti-IgG solid-phase peptide library screening [2] (1) Dissolve lyophilized IgG at a 5 g/L concentration in a solution of 50 mM sodium phosphate, 20 mM sodium chloride, and pH 8.3 medium. (2) Mix 1 mL of the (1) solution with 100 µL of additional dry DMF to dissolve 1 mg of AF488 NHS ester. Then, incubate for 1 hour at room temperature while rotating. (3) Use a 3-kDa MWCO filter in an Amicon Ultra 0.5-mL centrifugal filter unit to gather samples. (4) Using a pH 7.4 (50 mM sodium phosphate, 150 mM sodium chloride) solution at 5× precipitation resin volume, the deprotected hexamer or tetramer library was rinsed three times to reach equilibrium. (5) Reduce the concentration of IgG-AF488 to 1.3 mg/mL by diluting it with a solution of 50 mM sodium phosphate, 150 mM sodium chloride, and 0.2% Tween (pH 7.4). At 2–8°C, incubate (4) and (5) for the entire night. 7.4 (PBS-T) with 50 mM sodium phosphate, 150 mM sodium chloride, and 0.1% Tween 20 is used to wash the resin beads. (8) Using a fluorescence microscope set to 10x magnification, one bead of resin was placed in each well of a 96-well plate containing 40 µL PBS-T. The threshold for fluorescence screening and measurement with Alexa Fluor 488 under 480 nm excitation was set at 510 nm emission intensity. 20 mM AF488 NHS ester should be prepared using DMF. AF488 NHS ester stock solution [1]. It is advised to store the AF488 NHS ester stock solution at -20°C in the dark following aliquoting.
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References |
[1]. Kendall CG, et al. Amine Analysis Using AlexaFluor 488 Succinimidyl Ester and Capillary Electrophoresis with Laser-Induced Fluorescence. J Anal Methods Chem. 2015;2015:368362.
[2]. Lavoie RA, et al. Targeted Capture of Chinese Hamster Ovary Host Cell Proteins: Peptide Ligand Discovery. Int J Mol Sci. 2019 Apr 8;20(7):1729. |
Molecular Formula |
C25H17N3O13S2
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Molecular Weight |
631.54
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CAS # |
1374019-99-4
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Related CAS # |
AF488 NHS ester TEA
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Appearance |
Typically exists as solids (or liquids in special cases) at room temperature
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SMILES |
C(C1C=C(C(=O)ON2C(CCC2=O)=O)C=CC=1C1=C2C=CC(N)=C(S(O)(=O)=O)C2=[O+]C2C(=C(N)C=CC1=2)S([O-])(=O)=O)(=O)O
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HS Tariff Code |
2934.99.9001
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Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80: Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 :Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400  (Please use freshly prepared in vivo formulations for optimal results.) |
Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
1 mM | 1.5834 mL | 7.9172 mL | 15.8343 mL | |
5 mM | 0.3167 mL | 1.5834 mL | 3.1669 mL | |
10 mM | 0.1583 mL | 0.7917 mL | 1.5834 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration: mg/mL;
Method for preparing DMSO stock solution: mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.