Trypanosoma brucei rhodesiense Stephens and Fantham

Trypanosoma brucei rhodesiense Stephens and Fantham

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货期:

编号:TS205030

品牌:Testobio

产品名称：	Trypanosoma brucei rhodesiense Stephens and Fantham
商品货号：	TS205030
Deposited As：	Trypanosoma brucei rhodesiense
Strain Designations：	KETRI 269
Biosafety Level：	2 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Isolation：	Human clinical isolate, Kitanga, Tanzania, 1960
Product Format：	frozen
Storage Conditions：	Frozen Cultures: -70°C for 1 week; liquid N₂ vapor for long term storage Freeze-dried Cultures: 2-8°C Live Cultures: See Protocols section for handling information
Comments：	Produces acute and fatal parasitemia in mice
Growth Conditions：	Culture System: In vivo, Balb/c mouse
Cryopreservation：	Reagents Trypanosome Dilution Buffer 20 mM Na₂HPO₄ 2 mM NaH₂PO₄ 80 mM NaCl 5 mM KCl 1 mM MgSO₄ 20 mM glucose Adjust the pH of the solution to 7.7 and filter sterilize. Harvest and Preservation Prepare a 40% (v/v) sterile glycerol solution in Trypanosome Dilution Buffer (TDB). Dispense 0.5 mL of anticoagulant solution into a 15 mL test tube. Add to the anticoagulant blood collected by orbital bleeding from mice that had reached or are near peak parasitemia. Invert the tube several times to mix the blood with the anticoagulant.xa0 In a separate test tube, add the heparinized blood dropwise to the 40% glycerol solution. Note that blood should be mixed with glycerol solution in a 1:1 ratio to obtain a final concentration of cryoprotectant of 20% (v/v).xa0 Mix slowly by inversion and place the tube on ice.xa0 The freezing process should start 15 to 30 minutes following the addition of the heparinized blood to the cryoprotectant solution. Dispense 0.5 mL aliquots of blood suspension into 1.0 - 2.0 mL sterile plastic screw-capped cryovials.xa0 Place the vials in a controlled rate freezing unit.xa0 From room temperature cool the vials at -1°C/min to -40°C.xa0 If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through this phase.xa0 At -40°C, plunge vials into liquid nitrogen. Alternatively, place the vials in a Nalgene 1^oC freezing container.xa0 Place the container at -80°C for 1.5 to 2 hours and then plunge vials into liquid nitrogen. To thaw a frozen ampule, place in a 35°C water bath, until thawed (2-3 min).xa0 Immerse the ampule just sufficient to cover the frozen material.xa0 Do not agitate the ampule. Immediately after thawing, aseptically remove the contents of the ampule with a syringe and inoculate a Balb/c mouse.xa0 Follow the protocol for maintenance of the culture above. xa0
Name of Depositor：	CJ Bacchi
References：	Bacchi CJ, et al. Differential susceptibility to DL-alpha-difluoromethylornithine in clinical isolates of Trypanosoma brucei rhodesiense. Antimicrob Agents Chemother 34(6): 1183-1188, 1990. PubMed: 2118325

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Trypanosoma brucei rhodesiense Stephens and Fantham

货号: TS205030
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产品名称：	Trypanosoma brucei rhodesiense Stephens and Fantham
商品货号：	TS205030
Deposited As：	Trypanosoma brucei rhodesiense
Strain Designations：	KETRI 269
Biosafety Level：	2 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Isolation：	Human clinical isolate, Kitanga, Tanzania, 1960
Product Format：	frozen
Storage Conditions：	Frozen Cultures: -70°C for 1 week; liquid N₂ vapor for long term storage Freeze-dried Cultures: 2-8°C Live Cultures: See Protocols section for handling information
Comments：	Produces acute and fatal parasitemia in mice
Growth Conditions：	Culture System: In vivo, Balb/c mouse
Cryopreservation：	Reagents Trypanosome Dilution Buffer 20 mM Na₂HPO₄ 2 mM NaH₂PO₄ 80 mM NaCl 5 mM KCl 1 mM MgSO₄ 20 mM glucose Adjust the pH of the solution to 7.7 and filter sterilize. Harvest and Preservation Prepare a 40% (v/v) sterile glycerol solution in Trypanosome Dilution Buffer (TDB). Dispense 0.5 mL of anticoagulant solution into a 15 mL test tube. Add to the anticoagulant blood collected by orbital bleeding from mice that had reached or are near peak parasitemia. Invert the tube several times to mix the blood with the anticoagulant.xa0 In a separate test tube, add the heparinized blood dropwise to the 40% glycerol solution. Note that blood should be mixed with glycerol solution in a 1:1 ratio to obtain a final concentration of cryoprotectant of 20% (v/v).xa0 Mix slowly by inversion and place the tube on ice.xa0 The freezing process should start 15 to 30 minutes following the addition of the heparinized blood to the cryoprotectant solution. Dispense 0.5 mL aliquots of blood suspension into 1.0 - 2.0 mL sterile plastic screw-capped cryovials.xa0 Place the vials in a controlled rate freezing unit.xa0 From room temperature cool the vials at -1°C/min to -40°C.xa0 If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through this phase.xa0 At -40°C, plunge vials into liquid nitrogen. Alternatively, place the vials in a Nalgene 1^oC freezing container.xa0 Place the container at -80°C for 1.5 to 2 hours and then plunge vials into liquid nitrogen. To thaw a frozen ampule, place in a 35°C water bath, until thawed (2-3 min).xa0 Immerse the ampule just sufficient to cover the frozen material.xa0 Do not agitate the ampule. Immediately after thawing, aseptically remove the contents of the ampule with a syringe and inoculate a Balb/c mouse.xa0 Follow the protocol for maintenance of the culture above. xa0
Name of Depositor：	CJ Bacchi
References：	Bacchi CJ, et al. Differential susceptibility to DL-alpha-difluoromethylornithine in clinical isolates of Trypanosoma brucei rhodesiense. Antimicrob Agents Chemother 34(6): 1183-1188, 1990. PubMed: 2118325

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