composicion de la solucion tl- hepes como extensor

TLHEPES Component M.W. mM amount/100 ml amount/500 ml 1. CaCl 2 .2H 2 O 147.02 2.0 30.0 mg 150 mg 2. KCl 74.56 3.2 23.5 mg 118 mg 3. MgCl 2 .6H 2 O 203.3 0.5 10 mg 50 mg 4. NaH 2 PO 4 .H 2 O 137.99 0.4 5.6 mg 28 mg 5. Lactic Acid (Sodium salt; 60% w/w syrup) 112.1 10.0 186 mg (141 µl) 930 mg (705 µl) 6. NaCl 58.44 114.0 666 mg 3330 mg 7. NaHCO 3 84.01 2.0 16.8 mg 84 mg 8. HEPES (Free Acid) 238.3 10.0 238 mg 1190 mg 9. Na Pyruvate (from stock ) 110.0 0.2 1.0 ml 5.0 ml 10. BSA (Fraction V) 3 mg/ml 300 mg 1500 mg 11. Gentamycin (from stock ) 50 µg/ml 100 µl 500 µl Add approximately 80% of the double distilled water need to a beaker. With stirring, add components 1 8 in order. Adjust pH to 7.4 at 2022 C using 1 or 2 N NaOH. Add water to bring to 100 % of volume. This can be stored up to two weeks as a stock at 5 C if filtered and kept sterile. Generally we make up 500 ml at a time. On day of use, add components 9 and 10. Note that these can be for any volume desired for use that day and need not be 100 or 500 ml. Filter to sterilize with 0.2 µm filters. See Stock solutions for components 9 and 11. Return to Media List

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composicion de Tl hepes un extensor para el manejo de espermatozoides asi como manejo de la muestra de espermatozoides para realizar estudios de infertilidad o el analisis normal delespermatozoide ya que brinda un mayor plazo de tiempo para poder trabajar la muestra

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4/15/2015 TLHepes

http://www.ansci.wisc.edu/jjp1/ansci_repro/lab/procedures/media/tl_hepes.html 1/1

TLHEPES

Component M.W. mM amount/100mlamount/500

ml1.CaCl2.2H2O 147.02 2.0 30.0mg 150mg2.KCl 74.56 3.2 23.5mg 118mg3.MgCl2.6H2O 203.3 0.5 10mg 50mg4.NaH2PO4.H2O 137.99 0.4 5.6mg 28mg5.LacticAcid(Sodiumsalt60%w/wsyrup)

112.1 10.0 186mg(141l)

930mg(705l)

6.NaCl 58.44 114.0 666mg 3330mg7.NaHCO3 84.01 2.0 16.8mg 84mg8.HEPES(FreeAcid) 238.3 10.0 238mg 1190mg9.NaPyruvate(fromstock) 110.0 0.2 1.0ml 5.0ml

10.BSA(FractionV) 3mg/ml 300mg 1500mg11.Gentamycin(fromstock) 50g/ml 100l 500l

Addapproximately80%ofthedoubledistilledwaterneedtoabeaker.Withstirring,addcomponents18inorder.AdjustpHto7.4at2022Cusing1or2NNaOH.Addwatertobringto100%ofvolume.Thiscanbestoreduptotwoweeksasastockat5Ciffilteredandkeptsterile.Generallywemakeup500mlatatime.Ondayofuse,addcomponents9and10.Notethatthesecanbeforanyvolumedesiredforusethatdayandneednotbe100or500ml.Filtertosterilizewith0.2mfilters.SeeStocksolutionsforcomponents9and11.

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