计算溶液所需的质量、体积或浓度。
Tris-氯化铵红细胞裂解液 (无菌)
有货
为什么选择此级别
BioReagent,无菌,无菌过滤 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
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2-8°C储存。低温运输 。请查阅批次 COA 获取详细规格。
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文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
去除红细胞的方法有多种，Tris-氯化铵红细胞裂解液 (Tris-NH₄Cl Red Blood Cell Lysis Buffer)，也称为Tris-NH₄Cl Lysis Buffer，是一种从人、鼠或其他哺乳动物等体内的组织样品或血液中裂解并去除无核红细胞的溶液，其主要有效成分为氯化铵，Tris-NH₄Cl Lysis Buffer 经过优化配方，在裂解无核红细胞的同时几乎不损伤淋巴细胞 (lymphocyte)或其它有细胞核的细胞。
Tris-NH₄Cl Lysis Buffer对于裂解、去除有细胞核红细胞，例如鸟或禽类的红细胞，效果不佳，裂解类似细胞时，不建议采用。本裂解液经过滤除菌，经过Tris-NH₄Cl Lysis Buffer 处理过的血液或组织细胞样品可以用于后续的细胞培养、细胞融合以及核酸或蛋白的提取及各种常规的分析和检测。
使用方法
(一) 组织细胞样本的常规操作
1、制备细胞悬液: 新鲜组织经过胰蛋白酶或胶原酶等消化处理，通过适当方法制备成细胞悬液，离心弃上清。
2、裂解: 加入3-5倍细胞沉淀体积的Tris-NH₄Cl Lysis Buffer，轻柔吹打混匀，裂解2-3 min。本操作步骤在 4℃条件下操作更佳，亦可在室温下操作。
3、离心: 将细胞置于100%的胎牛血清中，4℃，300-400g离心10 min，弃红色上清。如无低温离心机本步骤亦可在室温下操作。
4、如果发现红细胞裂解不完全，可以重复上述步骤2和步骤3各一次。
5、洗涤：根据具体实验要求加入适量 PBS、HBSS、生理盐水或无血清培养液，轻柔混匀重悬沉淀。4℃，300-400g离心3-5 min，弃上清，离心步骤亦可在室温下操作。所加入的PBS、HBSS、生理盐水或无血清培养液的量一般应大于细胞沉淀体积的5倍以上。
6、如有必要，重复上述步骤5一次，共洗涤1-2次。
7、根据实验需要用适当溶液重悬细胞沉淀，进行计数、培养等后续实验
(二) 组织细胞样本的快速操作 (无需洗涤)
1、制备细胞悬液：新鲜组织经过胰蛋白酶或胶原酶等消化处理，通过适当方法制备成细胞悬液，离心弃上清。
2、裂解：加入细胞5倍细胞沉淀体积的Tris-NH₄Cl Lysis Buffer，轻柔吹打混匀，裂解2-3 min。本操作步骤在4℃条件下操作更佳，亦可在室温下操作。
3、 加入15-20 mL的PBS、HBSS、生理盐水或无血清培养液，轻柔混匀。
4、 离心: 将细胞置于100%的胎牛血清中，4℃，300-400g离心5 min，弃红色上清，本步骤亦可在室温下操作。
5、如果发现红细胞裂解不完全，可以重复上述步骤2-4各一次。
6、根据实验需要用适当溶液重悬细胞沉淀，进行计数、培养等后续实验。
(三) 血液样本的常规操作
1、取新鲜抗凝血，400-500g离心5 min，离心弃上清。
2、裂解: 加入6-10倍细胞沉淀体积的Tris-NH₄Cl Lysis Buffer，轻柔吹打混匀，裂解2-5 min。本操作步骤在 4℃条件下操作更佳，亦可在室温下操作。（特别提醒: 对于鼠的血液，裂解 2-3 min已经足够，对于人的外周血，宜延长裂解时间至4-5 min，并且裂解过程中轻轻摇动以促进红细胞裂解。）
3、离心: 将细胞置于100%的胎牛血清中，4℃，300-400g离心10 min，弃红色上清。如无低温离心机本步骤亦可在室温下操作。
4、如果发现红细胞裂解不完全，可以重复上述步骤2和步骤3一次。
5、洗涤: 根据具体实验要求加入适量PBS、HBSS、生理盐水或无血清培养液，轻柔混匀重悬沉淀。4℃，300-400g离心3-5 min，弃上清，离心步骤亦可在室温下操作。所加入的PBS、HBSS、生理盐水或无血清培养液的量一般应大于细胞沉淀体积的5倍以上。
6、根据实验需要用适当溶液重悬细胞沉淀，进行计数、培养等后续实验。 注意: 对于微量或少量的血液样本，可以不用第1步操作，加入10倍血液体积的Tris-NH₄Cl Lysis Buffer进行第2步操作，并在4℃或室温裂解4-15 min。对于鼠的血液，裂解4-5 min已经足够；对于人的外周血，宜延长裂解时间至10 min，但通常不宜超过15 min，并且裂解过程中宜适当摇动以促进红细胞裂解。
(四) 血液样本的快速操作 (无需洗涤)
1、新鲜抗凝血中加入10倍体积的Tris-NH₄Cl Lysis Buffer，轻轻吹打混匀， 裂解4-15 min。本操作步骤在 4℃条件下操作更佳，亦可在室温下操作。（特别提醒: 对于鼠的血液，裂解4-5min已经足够，对于人的外周血，宜延长裂解时间至10 min，但通常不宜超过15 min，并且裂解过程中宜适当摇动以促进红细胞裂解。
2、加入20-30 mL PBS、HBSS、生理盐水或无血清培养液，轻柔混匀。
3、将细胞置于100%的胎牛血清中，300-400g离心10 min，弃红色上清。4℃离心效果更佳。
4、如果发现红细胞裂解不完全，可以重复上述步骤2和步骤3一次。
5、 根据实验需要用适当溶液重悬细胞沉淀，进行计数、培养等后续实验。
注意事项
1、制备细胞悬液是应根据具体实验需要，不一定要制备成单细胞悬液。
2、后续试验如果是用于细胞培养，操作过程中应注意无菌操作，尽量在超净工作台内操作。
3、离心步骤尽量在4℃离心机上操作。
4、对于常规步骤与快速步骤的区别在于：常规步骤多了一步洗涤过程的离心，可以节省洗涤液的用量，并且洗涤效果也更好，不需要大体积的离心管；快速步骤少了一次离心过程，洗涤效果略差一些，同时需要大体积的离心管。
5、离心洗涤后，通常极微量的红细胞不会影响后续的检测。
6、如果经过Tris-NH₄Cl Lysis Buffer处理后的样品后续用于总RNA的提取，在处理细胞时不必使用DEPC处理的溶液，即无需在该操作中特意去除RNase。
7、为了您的安全和健康，请穿实验服并戴一次性手套操作。
8、本产品仅用于科研实验，禁止他用。
Product Description
There are various methods for removing red blood cells. Tris-Ammonium Chloride Red Blood Cell Lysis Buffer (Tris-NH₄Cl Red Blood Cell Lysis Buffer), also referred to as Tris-NH₄Cl Lysis Buffer, is a solution designed to lyse and eliminate enucleated red blood cells from tissue samples or blood of humans, mice, or other mammals. Its main active component is ammonium chloride. With an optimized formulation, Tris-NH₄Cl Lysis Buffer can lyse enucleated red blood cells while causing almost no damage to lymphocytes or other nucleated cells.
This lysis buffer has poor efficacy in lysing and removing nucleated red blood cells, such as those from birds or avian species, and is not recommended for use with such cells. The lysis buffer is sterilized by filtration. Blood or tissue cell samples treated with Tris-NH₄Cl Lysis Buffer can be used for subsequent experiments including cell culture, cell fusion, nucleic acid or protein extraction, as well as various routine analyses and detections.
Operating Procedures
(1) Routine Operation for Tissue Cell Samples
1. Prepare cell suspension: Digest fresh tissues with trypsin, collagenase, or other suitable enzymes, prepare a cell suspension using an appropriate method, and centrifuge to discard the supernatant.
2. Lysis: Add 3-5 volumes of Tris-NH₄Cl Lysis Buffer relative to the cell pellet volume, gently pipette to mix thoroughly, and incubate for lysis at 2-3 minutes. This step is optimally performed at 4℃, but can also be carried out at room temperature.
3. Centrifugation: Resuspend the cells in 100% fetal bovine serum, centrifuge at 300-400×g for 10 minutes at 4℃, and discard the red supernatant. If a low-temperature centrifuge is unavailable, this step can be performed at room temperature.
4. If red blood cell lysis is incomplete, repeat steps 2 and 3 once.
5. Washing: Add an appropriate volume of PBS, HBSS, normal saline, or serum-free medium according to specific experimental requirements, gently mix to resuspend the pellet. Centrifuge at 300-400×g for 3-5 minutes at 4℃, and discard the supernatant. This centrifugation step can also be performed at room temperature. The volume of added PBS, HBSS, normal saline, or serum-free medium should generally be more than 5 times the volume of the cell pellet.
6. If necessary, repeat step 5 once, with a total of 1-2 washing cycles.
7. Resuspend the cell pellet with an appropriate solution according to experimental needs, and proceed with subsequent experiments such as cell counting and culture.
(2) Rapid Operation for Tissue Cell Samples (No Washing Required)
1. Prepare cell suspension: Digest fresh tissues with trypsin, collagenase, or other suitable enzymes, prepare a cell suspension using an appropriate method, and centrifuge to discard the supernatant.
2. Lysis: Add 5 volumes of Tris-NH₄Cl Lysis Buffer relative to the cell pellet volume, gently pipette to mix thoroughly, and incubate for lysis at 2-3 minutes. This step is optimally performed at 4℃, but can also be carried out at room temperature.
3. Add 15-20 mL of PBS, HBSS, normal saline, or serum-free medium, and mix gently.
4. Centrifugation: Resuspend the cells in 100% fetal bovine serum, centrifuge at 300-400×g for 5 minutes at 4℃, and discard the red supernatant. This step can also be performed at room temperature.
5. If red blood cell lysis is incomplete, repeat steps 2-4 once.
6. Resuspend the cell pellet with an appropriate solution according to experimental needs, and proceed with subsequent experiments such as cell counting and culture.
(3) Routine Operation for Blood Samples
1. Take fresh anticoagulated blood, centrifuge at 400-500×g for 5 minutes, and discard the supernatant.
2. Lysis: Add 6-10 volumes of Tris-NH₄Cl Lysis Buffer relative to the cell pellet volume, gently pipette to mix thoroughly, and incubate for lysis at 2-5 minutes. This step is optimally performed at 4℃, but can also be carried out at room temperature. (Important Note: For mouse blood, 2-3 minutes of lysis is sufficient; for human peripheral blood, it is advisable to extend the lysis time to 4-5 minutes with gentle shaking during lysis to facilitate red blood cell lysis.)
3. Centrifugation: Resuspend the cells in 100% fetal bovine serum, centrifuge at 300-400×g for 10 minutes at 4℃, and discard the red supernatant. If a low-temperature centrifuge is unavailable, this step can be performed at room temperature.
4. If red blood cell lysis is incomplete, repeat steps 2 and 3 once.
5. Washing: Add an appropriate volume of PBS, HBSS, normal saline, or serum-free medium according to specific experimental requirements, gently mix to resuspend the pellet. Centrifuge at 300-400×g for 3-5 minutes at 4℃, and discard the supernatant. This centrifugation step can also be performed at room temperature. The volume of added PBS, HBSS, normal saline, or serum-free medium should generally be more than 5 times the volume of the cell pellet.
6. Resuspend the cell pellet with an appropriate solution according to experimental needs, and proceed with subsequent experiments such as cell counting and culture. Note: For trace or small-volume blood samples, step 1 can be omitted. Add 10 volumes of Tris-NH₄Cl Lysis Buffer relative to the blood volume and perform step 2, incubating for lysis at 4℃ or room temperature for 4–15 minutes. For mouse blood, 4-5 minutes of lysis is sufficient; for human peripheral blood, it is advisable to extend the lysis time to 10 minutes (usually not exceeding 15 minutes) with appropriate shaking during lysis to facilitate red blood cell lysis.
(4) Rapid Operation for Blood Samples (No Washing Required)
1. Add 10 volumes of Tris-NH₄Cl Lysis Buffer to fresh anticoagulated blood, gently pipette to mix thoroughly, and incubate for lysis at 4℃ or room temperature for 4-15 minutes. (Important Note: For mouse blood, 4-5 minutes of lysis is sufficient; for human peripheral blood, it is advisable to extend the lysis time to 10 minutes (usually not exceeding 15 minutes) with appropriate shaking during lysis to facilitate red blood cell lysis.)
2. Add 20-30 mL of PBS, HBSS, normal saline, or serum-free medium, and mix gently.
3. Resuspend the cells in 100% fetal bovine serum, centrifuge at 300-400×g for 10 minutes (centrifugation at 4℃ yields better results), and discard the red supernatant.
4. If red blood cell lysis is incomplete, repeat steps 2 and 3 once.
5. Resuspend the cell pellet with an appropriate solution according to experimental needs, and proceed with subsequent experiments such as cell counting and culture.
Precautions
1. When preparing the cell suspension, it is not necessary to prepare a single-cell suspension unless required by specific experiments.
2. If subsequent experiments involve cell culture, aseptic techniques should be strictly followed, and operations should be performed in a clean bench as much as possible.
3. Centrifugation steps are preferably carried out using a low-temperature centrifuge set at 4℃.
4. The difference between routine and rapid procedures lies in that the routine procedure includes an additional centrifugation step for washing, which saves washing buffer consumption, achieves better washing efficiency, and does not require large-volume centrifuge tubes. In contrast, the rapid procedure omits one centrifugation step, resulting in slightly reduced washing efficiency and requiring large-volume centrifuge tubes.
5. After centrifugation and washing, trace amounts of residual red blood cells generally do not interfere with subsequent detections.
6. If samples treated with Tris-NH₄Cl Lysis Buffer are to be used for total RNA extraction, there is no need to use DEPC-treated solutions during the operation, i.e., RNase removal is not required in this process.
7. For your safety and health, wear a lab coat and disposable gloves during operation.
8. This product is for research use only and is prohibited for any other purposes.
规格
别名
Tris-氯化铵红细胞裂解液 | Tris-氯化铵裂解液
英文别名
Tris-Ammonium Chloride Red Blood Cell Lysis Buffer | Tris-NH₄Cl Red Blood Cell Lysis Buffer | Tris-NH₄Cl Lysis Buffer
规格或纯度
无菌过滤, BioReagent, 无菌
稳定性与储存
Store at 2-8℃ long term (12 months).
英文名称
Tris-Ammonium Chloride Red Blood Cell Lysis Buffer (Sterile)
储存条件
2-8°C储存
运输条件
低温运输
产品线
生物产品
名称和识别符
分子类型
生物试剂/缓冲液
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0333333 |
分析证书 | T1509568 |
| ZJ26F0333332 |
分析证书 | T1509568 |
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