概述
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产品名称Anti-Phospholipase A2抗体
参阅全部 Phospholipase A2 一抗 -
描述兔多克隆抗体to Phospholipase A2
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经测试应用适用于: IHC-P, WB, ICC/IFmore details
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种属反应性与反应: Human
预测可用于: Mouse, Rat, Rabbit, Horse, Cow, Dog, Orangutan
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免疫原
Synthetic peptide conjugated to KLH derived from within residues 700 to the C-terminus of Human Phospholipase A2.
(Peptide available as ab86259.)
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阳性对照
- This antibody gave a positive signal in HeLa Whole Cell Lysate - Bleomycin Treated (40U/ml)
性能
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形式Liquid
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存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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存储溶液Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
纯度Immunogen affinity purified
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克隆多克隆
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同种型IgG
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研究领域
相关产品
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Assay kits
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Compatible Secondaries
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Immunizing Peptide (Blocking)
应用
Our Abpromise guarantee covers the use of ab73406 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| 应用 | Ab评论 | 说明 |
|---|---|---|
| IHC-P | Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 85 kDa). | |
| ICC/IF | Use a concentration of 1 µg/ml. |
靶标
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功能Selectively hydrolyzes arachidonyl phospholipids in the sn-2 position releasing arachidonic acid. Together with its lysophospholipid activity, it is implicated in the initiation of the inflammatory response.
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组织特异性Expressed in various tissues such as macrophages, platelets, neutrophils, fibroblasts and lung endothelium.
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序列相似性Contains 1 C2 domain.
Contains 1 PLA2c domain. -
结构域The N-terminal C2 domain associates with lipid membranes upon calcium binding. It modulates enzyme activity by presenting the active site to its substrate in response to elevations of cytosolic Ca(2+).
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翻译后修饰Activated by phosphorylation at both Ser-505 and Ser-727.
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细胞定位Cytoplasm. Cytoplasmic vesicle. Translocates to membrane vesicles in a calcium-dependent fashion.
- Information by UniProt
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数据库链接
- Entrez Gene: 100033889 Horse
- Entrez Gene: 151056 Human
- Entrez Gene: 5321 Human
- Entrez Gene: 18783 Mouse
- Entrez Gene: 665270 Mouse
- Entrez Gene: 100173125 Orangutan
- Entrez Gene: 192259 Rat
- Omim: 600522 Human
see all -
别名
- Calcium dependent phospholipid binding protein antibody
- CPLA 2 antibody
- cPLA2 alpha antibody
see all
Anti-Phospholipase A2 antibody 图像
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Western blot - Anti-Phospholipase A2 antibody (ab73406)
Predicted band size : 85 kDaLane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Phospholipase A2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: A549 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab73406 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab73406 was shown to recognize Phospholipase A2 when Phospholipase A2 knockout samples were used, along with additional cross-reactive bands. Wild-type and Phospholipase A2 knockout samples were subjected to SDS-PAGE. Ab73406 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Phospholipase A2 antibody (ab73406)ICC/IF image of ab73406 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab73406, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa cells at 1µg/ml. -
Western blot - Phospholipase A2 antibody (ab73406)Anti-Phospholipase A2 antibody (ab73406) at 1 µg/ml + HeLa Whole Cell Lysate - Bleomycin Treated (40U/ml) at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 85 kDa
Observed band size : 100 kDa (why is the actual band size different from the predicted?)
The 100kDa band is comparable to molecular weights seen with other commercially available antibodies to Phospholipase A2 -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Phospholipase A2 antibody (ab73406)IHC image of Phospholipsae A2 staining in humam colon carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab73406, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
实验方案
Anti-Phospholipase A2 antibody (ab73406)参考文献
ab73406 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"