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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
RT，避光
- 保质期：
两年
- 英文名：
Mouse Bone Marrow Neutrophil Isolation Solution Kit
- 库存：
大量现货
- 供应商：
北京索莱宝科技有限公司
- 规格：
200ml/KIT
注意：本产品试剂组分和操作步骤发生变化，使用前请仔细阅读说明书。
保存：本产品对光敏感，应该室温避光储存，保质期 2 年。无菌开封后，保存于室温。
试剂盒组成：
试剂 A 200mL 室温避光
试剂 C 100mL 室温避光
红细胞沉降液 100mL 室温
细胞洗涤液 200mL 室温
操作步骤一： 红细胞沉降（仅供参考）
沉降去除红细胞：取骨髓单细胞悬液与血液稀释液(注射用生理盐水或 1×PBS)及红细胞沉降液按 照 1:1:1 比例混匀，室温下静置 30-40min 后，可见红细胞沉于试管底部，小心吸取上清层(富含白 细胞及血小板)用于后续细胞分离。
操作步骤二：
样本分离（仅供参考）
1. 样本分离：当样本体积小于 5mL 时,先在离心管中先加入 4mL 试剂 A，后将 2mL 试剂 C 小心叠加于试剂 A 之上，形成梯度界面(样本体 积大于等于 5mL，试剂 A 与试剂 C 比例 2:1，试剂总量等于沉降后样 本含量),将悬液小心平铺于分离液液面上方。由于密度差异，可见明 显的分层界面(注意二者的总体积不能超过离心管的三分之二，否则会 影响分离效果)。
2. 室温条件下水平转子 600-1000g ，25-30min(血液体积越大所需离 心力越大,离心时间越长,最佳分离条件需摸索，离心最大转速不超过 1000g)。
3. 离心后，离心管中可见两个环状乳白色层，上层为单个核细胞层,下层即所需中性粒细胞层(受 个体差异及分离条件影响，粒细胞白膜层可能会出现颜色较浅的情况)。
4. 用吸管吸取下层中性粒细胞白膜层至新的离心管中，加入 10mL 细胞清洗液洗涤细胞，250g，离 心 10min。
5. 弃上清,加入 5mL 细胞清洗液重悬细胞,250g，离心 5min。
6. 弃上清，重悬细胞备用。
骨髓单细胞悬液制备：
小动物骨髓的采集：
1. 处死动物，无菌提取股骨和胫骨，剪去两端软骨，露出红色的骨髓腔（注意尽可能少的剪走骨 髓腔）。
2. 取 1ml 的无菌注射器，吸取少量的含有 10%标准胎牛血清的稀释液或者是含有血清的培养基，冲 洗骨髓腔以获得骨髓。
3. 最终制备成 2×10 8–1×10 9 /ml 的单细胞悬液备用。
大动物骨髓的采集：
大动物骨髓的采集可采取活体穿刺方法：先将动物麻醉、固定、局部除毛、消毒皮肤，然后估计好 皮肤到骨髓的距离，把骨髓穿刺针的长度固定好。操作人员用左手把穿刺点周围的皮肤绷紧，右手 将穿刺针在穿刺点垂直刺入，轻轻左右旋转将穿刺针钻入，当穿刺针进入骨髓腔时常有落空感。连 接注射器缓慢抽吸骨髓组织，当注射器内抽到少许骨髓时即停止抽吸。用含 10%标准胎牛血清的稀 释液调整细胞浓度为 2×10 8–1×10 9 /ml 的单细胞悬液备用。
常用的骨髓穿刺点：
股骨：穿刺部位在股骨内侧面,靠下端的凹面处；
胸骨：穿刺部位是胸骨体与胸骨柄连接处；
肋骨：穿刺部位是第 5～7 肋骨各点的中点；
胫骨：穿刺部位是股骨内侧、靠下端的凹面处。
如果穿刺采用的是肋骨，穿刺结束后要用胶布封贴 穿刺孔，防止发生气胸。
注意事项：
A.开封前颠倒混匀，本分离液为无菌产品，为延长分离液保存时间，请在无菌条件下启封，避免 微生物污染。
B.分离液使用时应始终保持室温（18℃~25℃），如室内温度较低，可将分离液预热。4℃或者是 温度较低的条件下离心，可能会导致白膜层不清晰。
C.血液样本最好为新鲜抗凝血（采血 2h 以内），为保持中性粒细胞的活性，应避免冷冻和冷藏。
D.稀释血液或洗涤细胞，不可使用含 Ca、Mg 离子的缓冲液及培养液，其成分会导致血细胞凝集， 大大降低细胞得率及纯度。
E.部分塑料制品（如聚苯乙烯）因其带有的静电作用，可能会导致细胞挂壁，影响分离效果。
F.血液样本的粘稠度或者是温度差异，可能会影响分离效果，可以调节离心转数和离心时间，摸索 最佳的分离条件。
G.如果要进一步对分离的细胞进行培养，注意全程保持无菌操作，避免微生物污染。
H.不同动物血液在不同比重分离液中的细胞离散系数及细胞带电不同，用户在制定分离液时应提 供所需分离液的比重、动物种属及被分离细胞的名称。
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Deficiency of hasB accelerated the clearance of Streptococcus equi subsp. Zooepidemicus through gasdermin d-dependent neutrophil extracellular traps
|
AuthorShun Li, Guobin Xu, Zheng Guo, Yuxuan Liu, Zhiliang Ouyang, Yajuan Li, Yunfei Huang, Qinqin Sun, Bikash R. Giri, Qiang Fu |
Publish_toINTERNATIONAL IMMUNOPHARMACOLOGY |
|
IF4.8000 |
PMID39083933 |
LncRNA-mRNA co-expression analysis reveals aquaporin-9-promoted neutrophil extracellular trap formation and inflammatory activation in sepsis
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AuthorPingping Liang, Manman Zhu, Xingzi Sun, Li Wang, Bin Li, Siqi Ming, Muhammad Younis, Jianhua Yang, Yongjian Wu, Xi Huang |
Publish_toINTERNATIONAL IMMUNOPHARMACOLOGY |
|
IF4.8000 |
PMID39133961 |
5-oxoETE promote thrombosis in antiphospholipid syndrome by triggering NETs formation through PLC/PKC/ERK pathway
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AuthorSong Xiaodong, Chen Xufeng, Wang Dong, Bai Jie |
Publish_toINFLAMMATION RESEARCH |
|
IF4.8000 |
PMID39377801 |
Aerosol inhalation of rhIL-10 improves acute lung injury in mice by affecting pulmonary neutrophil phenotypes through neutrophil-platelet aggregates
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AuthorHuan Qin, Jiangang Wang, Luyuan Bai, Huiqin Ding, Hailing Ding, Fengyi Zhang, Yantao Han |
Publish_toINTERNATIONAL IMMUNOPHARMACOLOGY |
|
IF4.8000 |
PMID39778276 |
Composite Hydrogel Modulates Intrinsic Immune-Cascade Neovascularization for Ocular Surface Reconstruction after Corneal Chemical Injury
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AuthorJun Zhang, Kun Xi, Guohua Deng, Xi Zou, Peirong Lu |
Publish_toGels |
|
IF4.6000 |
PMID37754358 |
Neutrophil extracellular trap-related mechanisms in acne vulgaris inspire a novel treatment strategy with adipose-derived stem cells
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AuthorYu Honghao, Zhang Boyu, Zhan Yuanyuan, Yi Yi, Jiang Qiong, Zhang Qi, Wu Yiping, Wu Min |
Publish_toScientific Reports |
|
IF4.6000 |
PMID38233540 |
Rhaponticin Alleviates Collagen-induced Arthritis by Inhibiting NLRP3/GSDMD-mediated Neutrophil Extracellular Traps
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AuthorZhang Jingjing, Xie Xinyue, Shen Qinhao, Yuan Chenchen, Lu Guotao, Xiao Weiming, Gong Weijuan, Fu Xiaoyan, Feng Xuebing |
Publish_toINFLAMMATION |
|
IF4.5000 |
PMID39725843 |
Sodium butyrate restricts neutrophils migration and NETs formation through reducing macrophage-derived CXCL16 in calculous cholecystitis
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AuthorHongsuo Chen, Jing Wang, Qingyu Ji, Zhenyu Jiang |
Publish_toHeliyon |
|
IF4.0000 |
PMID38322881 |
Picroside II promotes HSC apoptosis and inhibits the cholestatic liver fibrosis in Mdr2−/− mice by polarizing M1 macrophages and balancing immune responses
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AuthorJIA Kexin, MA Zhi, ZHANG Yinhao, XIE Kaihong, LI Jianan, WU Jianzhi, QU Jiaorong, LI Fanghong, LI Xiaojiaoyang |
Publish_toChinese Journal of Natural Medicines |
|
IF4.0000 |
PMID39059831 |
Notch1 siRNA and AMD3100 Ameliorate Metabolic Dysfunction-Associated Steatotic Liver Disease
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AuthorChunli Zhu, Yiheng Cheng, Lei Yang, Yifu Lyu, Jingjing Li, Pengbo Zhao, Ying Zhu, Xiaofei Xin, Lifang Yin |
Publish_toBiomedicines |
|
IF3.9000 |
PMID40002899 |
Punicalin attenuates LPS-induced acute lung injury by inhibiting inflammatory cytokine production and MAPK/NF-κB signaling in mice
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AuthorWentao Ji, Xiaoting Zhang, Chao Sang, Huixian Wang, Keqian Zhou, Yan Zhang, Lulong Bo |
Publish_toHeliyon |
|
IF3.7760 |
PMID37101633 |
Comparison and Evaluation of Two Combination Modes of Angiotensin for Establishing Murine Aortic Dissection Models
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AuthorHe Wenhui, Yu Sanjiu, Li Haoyang, He Ping, Xiong Tiantian, Yan Chaojun, Zhang Jingyu, Chen Shan, Guo Mei, Tan Xu, Zhong Dan, Sun Jianbin, Xu Zhizhen, Cheng Wei, Li Jun |
Publish_toJournal of Cardiovascular Translational Research |
|
IF3.4000 |
PMID37749480 |
Activated NETosis of bone marrow neutrophils up-regulates macrophage osteoclastogenesis via cGAS-STING/AKT2 pathway to promote osteoporosis
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AuthorYutong Guo, Hanzhang Zhou, Yixiang Wang, Yan Gu |
Publish_toEXPERIMENTAL CELL RESEARCH |
|
IF3.3000 |
PMID39988126 |
Caspase-1 deficiency impairs neutrophils recruitment and bacterial clearance in Streptococcus equi ssp. zooepidemicus infected mice
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AuthorZihua Lin, Yi Xia, Jianfeng Guo, Guobin Xu, Yuxuan Liu, Yalin Yang, Honglin Xie, Yunfei Huang, Qiang Fu |
Publish_toVETERINARY MICROBIOLOGY |
|
IF3.2460 |
PMID35395546 |
Trichinella spiralis excretory/secretory products from adult worms inhibit NETosis and regulate the production of cytokines from neutrophils
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AuthorWang Jing, Tang Bin, You Xihuo, Cai Xuepeng, Jia Wanzhong, Liu Xiaolei, Liu Mingyuan, Jin Xuemin, Ding Jing |
Publish_toParasites & Vectors |
|
IF3.2000 |
PMID37864246 |
Abnormal adenosine metabolism of neutrophils inhibits airway inflammation and remodeling in asthma model induced by Aspergillus fumigatus
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AuthorLiu Ting-ting, Wang Yue-li, Zhang Zhi, Jia Li-xin, Zhang Jing, Zheng Shuai, Chen Zhi-hua, Shen Hua-hao, Piao Chun-mei, Du Jie |
Publish_toBMC Pulmonary Medicine |
|
IF3.1000 |
PMID37452319 |
A high concentration of neutrophil extracellular traps is observed in humans and mice suffering from endometriosis
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AuthorYuting Sun, Junhong Cai, Yanan Zhang, Shan Bao |
Publish_toJOURNAL OF REPRODUCTIVE IMMUNOLOGY |
|
IF2.9000 |
PMID39657366 |
The TGF-β/MMP9/RAGE axis induces sRAGE secretion by neutrophils and promotes oral carcinogenesis
|
AuthorXiaolin Chen, Ningyuan Wang, Chenyang Jing, Jianghan Li, Xuanhao Liu, Yuquan Yang, Tianhuan Song, Hao Jia |
Publish_toBiochemistry and Biophysics Reports |
|
IF2.7000 |
PMID38463640 |
LASP1 inhibits the formation of NETs and alleviates acute pancreatitis by stabilizing F-actin polymerization in neutrophils
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AuthorSiqin Zhang, Zhihao Wang, Yuyan Zhang, Xiaowu Dong, Qingtian Zhu, Chenchen Yuan, Guotao Lu, Weijuan Gong, Yawei Bi, Yaodong Wang |
Publish_toBIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS |
|
IF2.5000 |
PMID39708397 |
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