Extraction buffer solution, rabbit brain extraction solution, PT detection reagent and PT detection kit
A technology for detection kits and detection reagents, which is applied in the field of medical detection and can solve problems such as easy precipitation and poor stability of PT reagents
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Embodiment 1
[0038] (1) Preparation of extraction buffer
[0039] 0.3% trisodium citrate, 0.3% anhydrous sodium acetate, adjust pH to 6.4 with acetic acid.
[0040] (2) Obtaining Rabbit Brain Extract
[0041] Dissolve 4.5% rabbit brain powder in the prepared extraction buffer (in g / mL, the mass volume ratio of rabbit brain powder to extraction buffer is 4.5:100), incubate at 39°C for 1 h, shake every 15 min Homogenize once, collect the supernatant after centrifugation at 3000RPM, which is the rabbit brain extract.
[0042] (3) Addition of lyoprotectant and heparin antagonist
[0043] Lyoprotectant, heparin antagonist and calcium ions were added to the rabbit brain extract. The protective agent is 0.5% bovine serum albumin, 1% glycine, the heparin antagonist is 0.03mg / ml polybrene, and the calcium ion is 15mM calcium chloride.
[0044] (4) freeze-drying, reconstitute with purified water after freeze-drying.
Embodiment 2
[0046] (1) Preparation of extraction buffer
[0047] 0.1% trisodium citrate, 1% anhydrous sodium acetate, adjust pH to 6.0 with acetic acid.
[0048] (2) Obtaining Rabbit Brain Extract
[0049] Dissolve 3% rabbit brain powder with the prepared extraction buffer (in g / mL, the mass volume ratio of rabbit brain powder to extraction buffer is 3:100), incubate at 33°C for 1.5h, every 15min Shake once, centrifuge at 3000RPM and collect the supernatant, which is the rabbit brain extract.
[0050] (3) Addition of lyoprotectant and heparin antagonist
[0051]Lyoprotectant, heparin antagonist and calcium ions were added to the rabbit brain extract. The protective agent is 5% peptone, 10% alanine, the heparin antagonist is 0.01 mg / ml protamine, and the calcium ion is 10 mM calcium lactate.
[0052] (4) freeze-drying, reconstitute with purified water after freeze-drying.
Embodiment 3
[0054] (1) Preparation of extraction buffer
[0055] 1% trisodium citrate, 0.1% anhydrous sodium acetate, adjust pH to 7.0 with acetic acid.
[0056] (2) Obtaining Rabbit Brain Extract
[0057] Dissolve 5% rabbit brain powder with the prepared extraction buffer (in g / mL, the mass volume ratio of rabbit brain powder to extraction buffer is 5:100), incubate at 45°C for 0.5h, every 15min Shake once, centrifuge at 3000RPM and collect the supernatant, which is the rabbit brain extract.
[0058] (3) Addition of lyoprotectant and heparin antagonist
[0059] Lyoprotectant, heparin antagonist and calcium ions were added to the rabbit brain extract. The protective agent is 3% bovine serum albumin, 5% glycine, the heparin antagonist is 0.1mg / ml polybrene, and the calcium ion is 25mM calcium carbonate.
[0060] (4) freeze-drying, reconstitute with purified water after freeze-drying.
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