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Mouse HMGB-1(High mobility group protein B1) ELISA Kit

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  • sandwich-Ab-ELISA-Elabscience
  • sandwich-Ab-ELISA-Elabscience
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  • sandwich-Ab-ELISA-Elabscience
  • sandwich-Ab-ELISA-Elabscience
  • sandwich-Ab-ELISA-Elabscience
  • sandwich-Ab-ELISA-Elabscience
  • sandwich-Ab-ELISA-Elabscience
  • sandwich-Ab-ELISA-Elabscience
    产品货号:E-EL-M0676c

    中文名:小鼠高迁移率族蛋白B1(HMGB-1)酶联免疫吸附测定试剂盒

    别称:HMGB1, HMG1, HMG3, SBP-1, Amphoterin

    规格:
    • 96T
    • 48T
    • 96T*5
    数量:
    - +
    价格: ¥3000

    反应性: 小鼠

    检测范围: 15.63~1000 pg/mL

    灵敏度: 9.38 pg/mL

    用途

    该试剂盒用于体外定量检测Mouse 血清,血浆或其他生物体液中HMGB-1浓度

    检测原理

    本试剂盒采用双抗体夹心ELISA法。用抗小鼠HMGB-1抗体包被于酶标板上,实验时样品或标准品中的小鼠HMGB-1会与包被抗体结合,游离的成分被洗去。依次加入生物素化的抗小鼠HMGB-1抗体和辣根过氧化物酶标记的亲和素。抗小鼠HMGB-1抗体与结合在包被抗体上的小鼠HMGB-1结合、生物素与亲和素特异性结合而形成免疫复合物,游离的成分被洗去。加入显色底物(TMB),TMB在辣根过氧化物酶的催化下呈现蓝色,加终止液后变成黄色。用酶标仪在450nm波长处测OD值,HMGB-1浓度与OD450值之间呈正比,通过绘制标准曲线计算出样品中HMGB-1的浓度。

    反应类型 夹心法
    规格 96T
    反应时间 3.5h
    反应性 Mouse
    检测方法 Colormetric
    检测范围 15.63—1000 pg/mL
    灵敏度 9.38 pg/mL
    样本体积 100μL
    样本类型 血清,血浆或其他生物体液

    特异性

    可检测样本中的小鼠HMGB-1,且与其它相关蛋白无明显交叉反应。

    重复性

    板内,板间变异系数均<10%。

    典型数据

    由于实验操作条件的不同(如操作者、移液技术、洗板技术和稳定条件等),标准曲线的OD值会有所差异。以下数据和曲线仅供参考,实验者需根据自己的实验建立标准曲线。

    (pg/mL) O.D Average Corrected
    1000 2.453
    2.489
    2.471 2.405
    500 1.573
    1.591
    1.582 1.516
    250 0.889
    0.883
    0.886 0.82
    125 0.448
    0.454
    0.451 0.385
    62.5 0.279
    0.263
    0.271 0.205
    31.25 0.175
    0.157
    0.166 0.1
    15.63 0.115
    0.121
    0.118 0.052
    0 0.056
    0.076
    0.066 --

    精密度

    板内精密度:低浓度样本,中浓度样本和高浓度样本分别在1块板子上检测20次。
    板间精密度:低浓度样本,中浓度样本和高浓度样本分别在3块板子上检测20次。

    Intra-assay Precision Inter-assay Precision
    Sample 1 2 3 1 2 3
    n 20 20 20 20 20 20
    Mean
    (pg/mL)
    50.50 98.56 334.18 52.21 95.09 337.87
    Standard
    deviation
    2.65 4.51 17.78 3.53 5.30 13.18
    C V (%) 5.25 4.58 5.32 6.76 5.57 3.90

    回收率

    分别往5个不同样本中添加已知浓度的目标蛋白,做回收实验,得出回收率范围和平均回收率。

    Sample Type Range (%) Average Recovery (%)
    Serum (n=8) 92-105 99
    EDTA plasma (n=8) 88-103 94
    Cell culture media (n=8) 83-98 90

    线性

    分别往5个样本中添加已知浓度的目标蛋白,做回收实验,得出回收率范围及平均回收率。将5个样本分别稀释2倍,4倍,8倍,16倍做回收实验,得出回收率范围及平均回收率。

    Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
    1:2 Range (%) 91-104 96-110 87-97
    Average (%) 97 103 92
    1:4 Range (%) 90-104 84-98 85-98
    Average (%) 97 90 91
    1:8 Range (%) 90-101 84-96 82-95
    Average (%) 95 89 88
    1:16 Range (%) 90-106 81-95 82-94
    Average (%) 98 87 87

    试剂盒组成及保存

    未拆封的试剂盒可在2-8℃保存一周;如果一周以后才使用试剂盒,请拆开试剂盒按照下表中的条件分别保存各组分。

    中文名称 英文名称 规格 保存条件
    ELISA酶标板(可拆卸) Micro ELISA Plate(Dismountable) 8 孔×12 条 -20℃,可存放6个月
    冻干标准品 Reference Standard 2 支
    浓缩生物素化抗体(100×) Concentrated Biotinylated Detection Ab 1支 120 μL
    浓缩HRP酶结合物(100×) Concentrated HRP Conjugate 1支 120 μL -20℃(避光),可存放6个月
    标准品&样品稀释液 Reference Standard & Sample Diluent 1瓶 20 mL 2-8℃.可存放6个月
    生物素化抗体稀释液 Biotinylated Detection Ab Diluent 1瓶 14 mL
    酶结合物稀释液 HRP Conjugate Diluent 1瓶 14 mL
    浓缩洗涤液(25×) Concentrated Wash Buffer (25×) 1瓶 30 mL
    底物溶液(TMB) Substrate Reagent 1瓶 10 mL 2-8℃(避光)
    反应终止液 Stop Solution 1瓶 10 mL 2-8℃
    封板覆膜 Plate Sealer 5 张
    产品说明书 Product Description 1 份
    质检报告 Certificate of Analysis 1 份

    说明:所有试剂瓶盖须旋紧以防止蒸发和微生物的污染。
    试剂体积以实际发货版说明书为准。相关试剂在分装时会比标签上标明的体积稍多一些,请在使用时量取而非直接倒出。

    试验所需自备物品

    • 1. 酶标仪(450nm波长滤光片)
    • 2. 高精度移液器,EP管及一次性吸头:0.5-10 μL, 2-20 μL, 20-200 μL, 200-1000 μL
    • 3. 37℃恒温箱,双蒸水或去离子水
    • 4. 吸水纸

    操作步骤

    1. 对应板孔中加入100μL标准品工作液或样本,37℃孵育90分钟

    2. 弃掉板内液体后,立即加入100μL生物素化抗体工作液,37℃孵育60分钟

    3. 弃掉板内液体,洗板3次

    4. 每孔加入100μL HRP酶结合物工作液,37℃孵育30分钟,弃掉板内液体,洗板5次

    5. 每孔加入90μL底物溶液,37℃孵育15分钟左右

    6. 每孔加入50μL终止液

    7. 立即在450nm波长下读数,处理数据

    客户发表文献

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      物种: Mouse
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      Journal: ADVANCED FUNCTIONAL MATERIALS(2019)
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      Journal: ACS Nano(2021)
      DOI: 10.1021/acsnano.0c09681
      PMID: 33470095
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      物种: Mouse
    5. Mature dendritic cell-derived dendrosomes swallow oxaliplatin-loaded nanoparticles to boost immunogenic chemotherapy and tumor antigen-specific immunotherapy
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      DOI: 10.1016/j.bioactmat.2021.12.020
      PMID: 35386340
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      Journal: Nature Communications(2021)
      DOI: 10.1038/s41467-021-24564-0
      PMID: 34262026
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      DOI: 10.1016/j.biomaterials.2021.121130
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    9. Injectable Liquid Crystal Formation System for Reshaping Tumor Immunosuppressive Microenvironment to Boost Antitumor Immunity: Postoperative Chemoimmunotherapy
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      DOI: 10.1002/smll.202004905
      PMID: 33206460
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      物种: Mouse
    10. Enhancing cancer chemo-immunotherapy by biomimetic nanogel with tumor targeting capacity and rapid drug-releasing in tumor microenvironment
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      Journal: Acta Pharmaceutica Sinica B(2021)
      DOI: 10.1016/j.apsb.2021.11.004
      PMID: 35646526
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    11. Liraglutide abrogates nephrotoxic effects of chemotherapies
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      Journal: PHARMACOLOGICAL RESEARCH(2023)
      DOI: 10.1016/j.phrs.2023.106680
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    12. Doxorubicin-polyglycerol-nanodiamond conjugate is a cytostatic agent that evades chemoresistance and reverses cancer-induced immunosuppression in triple-negative breast cancer
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      DOI: 10.1186/s12951-019-0541-8
      PMID: 31623629
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      DOI: 10.1126/sciimmunol.aau6426
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    14. Laser/GSH-Activatable Oxaliplatin/Phthalocyanine-Based Coordination Polymer Nanoparticles Combining Chemophotodynamic Therapy to Improve Cancer Immunotherapy
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      Journal: ACS Applied Materials & Interfaces(2021)
      DOI: 10.1021/acsami.1c11327
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      物种: Mouse
    15. Bioactive Injectable Hydrogel Dressings for Bacteria-Infected Diabetic Wound Healing: A “Pull–Push” Approach
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      Journal: ACS Applied Materials & Interfaces(2022)
      DOI: 10.1021/acsami.2c04300
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    16. Systemic administration of polymersomal oncolytic peptide LTX-315 combining with CpG adjuvant and anti-PD-1 antibody boosts immunotherapy of melanoma
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      Journal: JOURNAL OF CONTROLLED RELEASE(2021)
      DOI: 10.1016/j.jconrel.2021.06.032
      PMID: 34174350
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      物种: Mouse
    17. PDTAC: Targeted Photodegradation of GPX4 Triggers Ferroptosis and Potent Antitumor Immunity
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      Journal: JOURNAL OF MEDICINAL CHEMISTRY(2022)
      DOI: 10.1021/acs.jmedchem.2c00855
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      样本类型: B16 cell
      物种: Mouse
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      Journal: OncoImmunology(2022)
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    20. Ferroptosis contribute to neonicotinoid imidacloprid-evoked pyroptosis by activating the HMGB1-RAGE/TLR4-NF-κB signaling pathway
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      DOI: 10.3390/ijms232113264
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    22. Liposomal remdesivir inhalation solution for targeted lung delivery as a novel therapeutic approach for COVID-19
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      DOI: 10.1016/j.ajps.2021.09.002
      PMID: 34703490
      样本类型: serum BALF,A549 cell
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    24. Retinal pigment epithelial cell necroptosis in response to sodium iodate
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      DOI: 10.1038/cddiscovery.2016.54
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    25. TNF‐α/HMGB1 inflammation signalling pathway regulates pyroptosis during liver failure and acute kidney injury
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      DOI: 10.1111/cpr.12829
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    26. Galectin-3 Deficiency Facilitates TNF-α-Dependent Hepatocyte Death and Liver Inflammation in MCMV Infection
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      DOI: 10.3389/fmicb.2019.00185
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    27. Osthole Blocks HMGB1 Release From the Nucleus and Confers Protective Effects Against Renal Ischemia-Reperfusion Injury.
      IF: 4.566
      Journal: Frontiers in Physiology(2021)
      DOI: 10.3389/fphys.2021.735425
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    28. Cigarette smoke‐induced HMGB1 translocation and release contribute to migration and NF‐κB activation through inducing autophagy in lung macrophages
      IF: 4.658
      Journal: JOURNAL OF CELLULAR AND MOLECULAR MEDICINE(2019)
      DOI: 10.1111/jcmm.14789
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      物种: Human,Mouse
    29. Hypothermic Oxygenated Machine Perfusion Alleviates Donation After Circulatory Death Liver Injury Through Regulating P-selectin-dependent and -independent Pathways in Mice
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    30. Structure-based design of glycyrrhetinic acid derivatives as potent anti-sepsis agents targeting high-mobility group box-1
      IF: 4.831
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      DOI: 10.1016/j.bioorg.2020.104461
      样本类型: Plasma,Cell culture supernatant
      物种: Mouse
    31. The receptor for advanced glycation end products mediates dysfunction of airway epithelial barrier in a lipopolysaccharides-induced murine acute lung injury model
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      样本类型: Bronchoalveolar Lavage Fluid
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    32. MiR-181-5p protects mice from sepsis via repressing HMGB1 in an experimental model
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      Journal: European Review for Medical and Pharmacological Sciences(2020)
      DOI: 10.26355/eurrev_202009_23063
      PMID: 33015817
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    33. SIRT1-mediated HMGB1 deacetylation suppresses sepsis-associated acute kidney injury
      IF: 3.164
      Journal: AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY(2018)
      DOI: 10.1152/ajprenal.00119.2018
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    34. High-Mobility Group Box 1 Disrupts Metabolic Function with Cigarette Smoke Exposure in a Ceramide-Dependent Manner
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    35. Honokiol Reduces Fungal Load, Toll-Like Receptor-2, and Inflammatory Cytokines in Aspergillus fumigatus Keratitis
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      DOI: 10.1167/iovs.61.4.48
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    36. Re-exposure to alarmin HMGB1 and cytokine IL-1 beta induces differential innate immune response generation in mouse brain
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      DOI: 10.1016/j.jneuroim.2021.577625
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      样本类型: brain
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      DOI: 10.1016/j.neuroscience.2021.05.003
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      样本类型: Cell culture supernatant,Tissue homogenate(Brain)
      物种: Rat,Mouse
    38. JAK2/STAT1-mediated HMGB1 translocation increases inflammation and cell death in a ventilator-induced lung injury model
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      Journal: LABORATORY INVESTIGATION(2019)
      DOI: 10.1038/s41374-019-0308-8
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      样本类型: Cell lysate,Bronchoalveolar Lavage Fluid,Tissue homogen
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      DOI: 10.1186/s13071-022-05283-x
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      物种: Mouse
    40. miR-129-5p alleviates LPS-induced acute kidney injury via targeting HMGB1/TLRs/NF-kappaB pathway
      IF: 3.943
      Journal: INTERNATIONAL IMMUNOPHARMACOLOGY(2020)
      DOI: 10.1016/j.intimp.2020.107016
      PMID: 33039954
      样本类型: Serum
      物种: Mouse
    41. Hesperidin ameliorates lipopolysaccharide-induced acute lung injury in mice by inhibiting HMGB1 release
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      Journal: INTERNATIONAL IMMUNOPHARMACOLOGY(2015)
      DOI: 10.1016/j.intimp.2015.02.022
      样本类型: Bronchoalveolar Lavage Fluid
      物种: Mouse
    42. Protective effects of hesperidin on concanavalin A-induced hepatic injury in mice
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      Journal: INTERNATIONAL IMMUNOPHARMACOLOGY(2014)
      DOI: 10.1016/j.intimp.2014.05.018
      样本类型: Serum
      物种: Mouse
    43. the Nrf2-ARE Signaling Pathway
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      DOI: 10.1007/s10753-018-0915-3
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    44. Anti-inflammatory effects of astaxanthin against fungal keratitis
      IF: 1.33
      Journal: International Journal of Ophthalmology(2020)
      DOI: 10.18240/ijo.2020.11.01
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      样本类型: Tissue homogenate(Corneas)
      物种: Mouse
    45. Protective Role of Grape Seed Proanthocyanidins Against Ccl4 Induced Acute Liver Injury in Mice
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      Journal: MEDICAL SCIENCE MONITOR(2016)
      DOI: 10.12659/MSM.895552
      样本类型: Serum
      物种: Mouse
    46. High mobility group box-1 (HMGB-1) and its receptors in the pathogenesis of malaria-associated acute lung injury/acute respiratory distress syndrome in a mouse model
      Journal: Heliyon(2021)
      DOI: 10.1016/j.heliyon.2021.e08589
      样本类型: lung tissue,plasma
      物种: Mouse
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    Q

    费*士Submitted [ Jul 27 2022 ]

    问: 细胞培养上清忘记离心了,检测实验组数据不太对,是因为杂质和细胞碎片没离心吗

    回复

    A

    adminSubmitted [ Jul 27 2022 ]

    回答: ELISA 的样本都是均质澄清的溶液,杂质和细胞碎片会影响抗原抗体的结合,导致检测结果不准确。

    A

    adminSubmitted [ Jul 27 2022 ]

    回答: 

    Q

    费*士Submitted [ Jul 27 2022 ]

    问: 细胞培养上清忘记离心了,检测实验组数据不太对,是因为杂质和细胞碎片没离心吗

    回复

    A

    adminSubmitted [ Jul 27 2022 ]

    回答: 是的,ELISA的样本是均质溶液,里面含有杂质和细胞碎片会影响抗原抗体的结合,会导致结果不准确。

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