ADAM8 抗体 (AA 145-493)
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- 抗原 See all ADAM8 抗体
- ADAM8 (ADAM Metallopeptidase Domain 8 (ADAM8))
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抗原表位
- AA 145-493
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适用
- 小鼠
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宿主
- 兔
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克隆类型
- 多克隆
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标记
- This ADAM8 antibody is un-conjugated
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应用范围
- Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), Immunocytochemistry (ICC)
- 原理
- Polyclonal Antibody to A Disintegrin And Metalloprotease 8 (ADAM8)
- 特异性
- The antibody is a rabbit polyclonal antibody raised against ADAM8. It has been selected for its ability to recognize ADAM8 in immunohistochemical staining and western blotting.
- 交叉反应
- 人, 大鼠
- 纯化方法
- Antigen-specific affinity chromatography followed by Protein A affinity chromatography
- 免疫原
- Recombinant A Disintegrin And Metalloprotease 8 (ADAM8) corresdonding to Glu145~Cys493 with N-terminal His Tag
- 亚型
- IgG
- Top Product
- Discover our top product ADAM8 Primary Antibody
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- 应用备注
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Western blotting: 0.5-5 μg/mL Immunohistochemistry: 5-20 μg/mL Immunocytochemistry: 5-20 μg/mL Optimal working dilutions must be determined by end user.
- 说明
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The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.
- 限制
- 仅限研究用
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- by
- Department of Neurosurgery, Philipps-University Marburg
- No.
- #104303
- 日期
- 2021.02.11
- 抗原
- ADAM8
- Lot Number
- PAA620Mu01
- Method validated
- Western Blotting
- Positive Control
Wildtype bone marrow derived mouse macrophages
- Negative Control
Adam8 knock-out bone marrow derived mouse macrophages
- Notes
Passed. ABIN1866534 specifically recognizes Adam8 in lysates of bone marrow derived mouse macrophages.
- Primary Antibody
- ABIN1866534
- Secondary Antibody
- ABIN269949
- Full Protocol
- Isolate and subsequently grow Adam8 wt or ko bone marrow derived mouse macrophages in DMEM (Gibco, 11965-092, lot 2258599) supplemented with FBS superior (Sigma Aldrich, S0615, lot 0001640840), Pen Strep (Gibco, 15140-122, lot 2240835), and Murine M-CSF (PeproTech, 315-02, lot 0419245) at 37°C and 5% CO2 in 2 mL on a 6 well plate to 70% confluency.
- Lyse 800,000 cells in 50 µL RIPA buffer (50 mM HEPES pH 7.4, 150 mM NaCl, 1% (v/v) NP-40, 0.5% (w/v) sodium deoxycholate, 0.1% (w/v) SDS, 10 mM phenanthrolin, 10 mM EDTA, phosphatase and proteinase inhibitor).
- Determine total protein content of the lysates using Pierce BCA Protein Assay Kit (Thermo Scientific, 23225, lot RH237553).
- Denature 10 µg of total protein for 5 min at 95 °C in 1x Laemmli Buffer and subsequently separate them on a denaturing 10% SDS PAGE gel (polyacrylamide, Tris-HCl pH 6.8 for the stacking gel and pH 8.8 for separating gel, water, ammonium persulfate, SDS, tetramethyldiamine in a Mini-PROTEAN Tetra System (BioRad) for 1.5 h at 120 V.
- Transfer proteins onto nitrocellulose membrane (GE Healthcare Life Science, 10600001, lot A29635973) with a Western blotting system for 70 min at 150 mA.
- Block the membrane with TBST containing 5% milk or 5% BSA for 1 h at RT.
- Incubation with primary
- rabbit anti-ADAM8 antibody (antibodies-online, ABIN1866534, PAA620Mu01) diluted 1:1,000 in TBST containing 5% milk or 5% BSA for over night at 4°C.
- rabbit-anti TUBB antibody (Novus Biologicals, NB600-936, lot J) diluted 1:2,000 in TBST containing 5% milk over night at 4°C.
- Wash membrane 3x for 10 min with TBST.
- Incubation with secondary Donkey Anti-Rabbit IgG H&L (HRP) (abcam, ab97064, lot GR3209122-1) diluted 1:5,000 in TBST containing 5% milk for 1h at RT.
- Wash membrane 3x for 10 min with TBST.
- Reveal protein bands using Western Bright Chemilumineszenz Substrat Sirius (Biozym, 541021, no 200331-75) on a Developing machine, Royal Intas ChemoCam Imager (Inta Science Imaging) exposure time 20 sec (TBST with 5% milk) or 3 sec (TBST with 5% BSA).
- Experimental Notes
ABIN1866534 reveals protein bands and 90 kDa and 110 kDa specifically in the Adam8 wildtype lysates. They are not present in the Adam8 ko lysates. And additional unspecific band between these two bands in visible in all lysates.
Blocking with TBST containing 5% milk gives clearer results that blocking with TBST containing 5% BSA.
生效 #104303 (Western Blotting)Validation ImagesFull Methods -
- 状态
- Liquid
- 浓度
- Lot specific
- 缓冲液
- 0.01M PBS, pH 7.4, containing 0.05 % Proclin-300, 50 % glycerol.
- 储存液
- ProClin
- 注意事项
- WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
- 注意事项
- Avoid repeated freeze-thaw cycles.
- 储存条件
- 4 °C,-20 °C
- 储存方法
- Store at 4°C for frequent use. Stored at -20°C in a manual defrost freezer for two year without detectable loss of activity. Avoid repeated freeze-thaw cycles.
- 有效期
- 24 months
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- 抗原
- ADAM8 (ADAM Metallopeptidase Domain 8 (ADAM8))
- 别名
- A Disintegrin And Metalloprotease 8 (ADAM8 产品)
- 别名
- CD156 antibody, MS2 antibody, CD156a antibody, E430039A18Rik antibody, adam8 antibody, reprolysin antibody, si:zc217g15.7 antibody, zgc:64059 antibody, ADAM8 antibody, RGD1566394 antibody, ADAM metallopeptidase domain 8 antibody, a disintegrin and metalloproteinase domain 8 antibody, a disintegrin and metallopeptidase domain 8 antibody, ADAM metallopeptidase domain 8a antibody, ADAM8 antibody, LOC100304737 antibody, Adam8 antibody, adam8a antibody, adam8 antibody
- 背景
- CD156-A, MS2, CD156a, Disintegrin and metalloproteinase domain-containing protein 8
- 途径
- Activation of Innate immune Response, M Phase
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