重组Anti-pan PDE4抗体[EPR25202-101] (ab300108)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25202-101] to PDE4
- Suitable for: WB, ICC/IF, Dot blot, IHC-Fr, IHC-P, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
概述
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产品名称
Anti-pan PDE4抗体[EPR25202-101]
参阅全部 PDE4 一抗 -
描述
兔单克隆抗体[EPR25202-101] to PDE4 -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, Dot blot, IHC-Fr, IHC-P, Flow Cyt (Intra)more details
不适用于: IP -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa whole cell lysate; mouse and rat cerebellum and brain tissue lysates. DB: human PDE4A/B/C/D fragment. IHC-P: Human, mouse, and rat cerebrum FFPE tissue sections. IHC-Fr: Rat and mouse cerebrum fresh frozen tissues. ICC/IF: HeLa cells, mouse and rat primary neurons. Flow Cyt (Intra): K-562 and HeLa cells.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR25202-101 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab300108于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
1/1000. Detects a band of approximately 60-100 kDa (predicted molecular weight: 91 kDa).
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ICC/IF |
1/100.
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Dot blot |
1/1000.
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IHC-Fr |
1/50.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/5000.
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说明 |
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WB
1/1000. Detects a band of approximately 60-100 kDa (predicted molecular weight: 91 kDa). |
ICC/IF
1/100. |
Dot blot
1/1000. |
IHC-Fr
1/50. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/5000. |
靶标
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相关性
The cAMP-specific Phosphodiesterase type 4 (PDE4) family is comprised of 4 genes (PDE4 A, B, C and D) with multiple splice variants generated by RNA splicing and use of alternate initiation sites. The cAMP-dependent protein kinase A modulates the enzyme activity of some PDE4 family members by phosphorylation / dephosphorylation mechanisms. Such short-term regulatory mechanisms are necessary for rapid changes in the cAMP levels in cells. PDE4 family selective inhibitor, rolipram inhibits all members of the PDE4 family. In rodents inhibition of PDE4 enzymes by rolipram attenuated short- and long-term memory impairment produced by the administration of scopolamine and MK801. Interaction of these enzymes with cellular scaffolding proteins plays important role in PDE4 compartmentalization and cAMP-dependent signaling. -
细胞定位
Cell Membrane -
数据库链接
- Entrez Gene: 5141 Human
- Entrez Gene: 5142 Human
- Entrez Gene: 5143 Human
- Entrez Gene: 5144 Human
- SwissProt: P27815 Human
- SwissProt: Q07343 Human
- SwissProt: Q08493 Human
- SwissProt: Q08499 Human
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别名
- PDE4A antibody
- PDE4B antibody
- PDE4C antibody
see all
图片
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All lanes : Anti-pan PDE4 antibody [EPR25202-101] (ab300108) at 1/1000 dilution
Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 3 : DU 145 (human prostate carcinoma epithelial cell) whole cell lysate
Lane 4 : Mouse brain tissue lysate
Lane 5 : Mouse cerebellum tissue lysate
Lane 6 : Rat brain tissue lysate
Lane 7 : Rat cerebellum tissue lysate
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 91 kDa
Observed band size: 60-100 kDa why is the actual band size different from the predicted?
Exposure time: 59 secondsab181602 was used as the Loading Control at 1/200000 dilution.
Blocking/Diluting buffer and concentration: 5% NFDM/TBS
Low expression: DU145 (PMID: 24518597), K562.
The molecular weight observed is consistent with what has been described in the literature (PMID:9880581, 15717866). -
Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling pan PDE4 with ab300108 at 1/500 (1.044 µg/ml) followed by a Leica DS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab300108 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is Leica DS9800 (Bond™ Polymer Refine Detection) kit.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum (fresh) tissue labeling pan PDE4 with ab300108 at 1/50 (10.44 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution (Green). Confocal image showing positive staining on mouse cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab300108 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling pan PDE4 with ab300108 at 1/50 (10.44 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (green). Confocal image showing cytoplasmic staining in HeLa cell line.
Low expression: K-562.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (red). The Nuclear counterstain was DAPI (blue).Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
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Dot blot analysis of pan PDE4 using ab300108 at 1/1000 (0.522 µg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.
Exposure time: 8 seconds
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Lane 1: His-tagged human PDE4A fragment
Lane 2: His-tagged human PDE4B fragment
Lane 3: GST-tagged human PDE4C fragment
Lane 4: His-tagged human PDE4D fragment -
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast, Left) HeLa (human cervix adenocarcinoma epithelial cell, Right) cells labeling pan PDE4 with ab300108 at 1/5000 dilution (0.01ug) (red) compared with a Rabbit monoclonal IgG (ab172730) (black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Low expression: K-562.
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Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling pan PDE4 with ab300108 at 1/5000 (0.104 µg/ml) followed by a Leica DS9800 (Bond™ Polymer Refine Detection) kit. Positive staining on mouse cerebrum. The section was incubated with ab300108 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is Leica DS9800 (Bond™ Polymer Refine Detection) kit was used at a ready to use dilution.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling pan PDE4 with ab300108 at 1/5000 (0.104 µg/ml) followed by a Leica DS9800 (Bond™ Polymer Refine Detection) kit. Positive staining on rat cerebrum. The section was incubated with ab300108 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is Leica DS9800 (Bond™ Polymer Refine Detection) kit used at a ready to use dilution.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum (fresh) tissue labeling pan PDE4 with ab300108 at 1/50 (10.44 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing positive staining on rat cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab300108 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron labeling pan PDE4 with ab300108 at 1/50 (10.44 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (green). Confocal image showing positive staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 (4µg/ml) dilution (red) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution. The Nuclear counterstain was DAPI (blue).-ve control: ab11267 at 1/500 (4μg/ml) followed by ab150081 at 1/1000 (2 ug/ml) dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron labeling pan PDE4 with ab300108 at 1/50 (10.44 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (green). Confocal image showing positive staining in rat primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 (4µg/ml) dilution (red) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution. The Nuclear counterstain was DAPI (blue).-ve control: ab11267 at 1/500 (4μg/ml) followed by ab150081 at 1/1000 (2 ug/ml) dilution.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (0)
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