Anti-CNPase 抗体 [11-5B] (ab6319)
Key features and details
- Mouse monoclonal [11-5B] to CNPase
- Suitable for: Flow Cyt, ICC, IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG1
製品の概要
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製品名
Anti-CNPase antibody [11-5B]
CNPase 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [11-5B] to CNPase -
由来種
Mouse -
アプリケーション
適用あり: Flow Cyt, ICC, IHC-P, WBmore details -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Sheep, Rabbit, Cow, Dog, Pig, Rhesus monkey非交差種: Chicken, Guinea pig
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免疫原
Full length native protein (purified) corresponding to Human CNPase.
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ポジティブ・コントロール
- WB: HeLa and Hap1 cell lysates; Human and Mouse Spinal Cord and Brain tissue lysates; Rat Brain tissue lysate. IHC-P: FFPE human cerebral cortex tissue sections. ICC: SKNSH cell line
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特記事項
This antibody clone is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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精製度
IgG fraction -
ポリ/モノ
モノクローナル -
クローン名
11-5B -
アイソタイプ
IgG1 -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
アプリケーション
Our Abpromise guarantee covers the use of ab6319 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt | Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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ICC | Use at an assay dependent concentration. | |
IHC-P | Use at an assay dependent concentration. | |
WB | Use a concentration of 5 µg/ml. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa). |
ターゲット情報
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配列類似性
Belongs to the cyclic nucleotide phosphodiesterase family. -
細胞内局在
Membrane. Melanosome. Firmly bound to membrane structures of brain white matter. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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参照データベース
- Entrez Gene: 280752 Cow
- Entrez Gene: 607694 Dog
- Entrez Gene: 1267 Human
- Entrez Gene: 12799 Mouse
- Entrez Gene: 493772 Pig
- Entrez Gene: 25275 Rat
- Entrez Gene: 641337 Rhesus monkey
- Omim: 123830 Human
see all -
別名
- 2'' antibody
- 2'3' cyclic nucleotide 3' phosphodiesterase antibody
- 3''-cyclic-nucleotide 3''-phosphodiesterase antibody
see all
画像
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All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 5 µg/ml
Lane 1 : Wild-type HAP1 cell lysate at 40 µg
Lane 2 : CNPase knockout HAP1 cell lysate at 40 µg
Lane 3 : Human brain whole cell lysate at 20 µg
Predicted band size: 48 kDaLanes 1 - 3: Merged signal (red and green). Green - ab6319 observed at 48 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab6319 was shown to recognize CNPase in wild-type HAP1 cells as signal was lost at the expected MW in CNPase knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CNPase knockout samples were subjected to SDS-PAGE. Ab6319 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 5 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNPase antibody [11-5B] (ab6319)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom
IHC-P image of CNPase staining on rat brain sections using ab6319 (1/1600). heat mediated antigen retrieval on paraffin embedded sections was performed using citric acid. The sections were then blocked with 1% BSA for 10 min at 21°C. The primary antibody was incubated for 16 hours at 21°C. The sections were then incubated in Goat anti-mouse (Biotin) at 1:200.
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All lanes : Anti-CNPase antibody [11-5B] (ab6319)
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CNP knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 48 kDa
Observed band size: 48 kDaLanes 1- 2: Merged signal (red and green). Green - ab6319 observed at 48 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.
ab6319 was shown to react with CNPase in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264949 (knockout cell lysate ab256877) was used. Wild-type HeLa and CNP knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab6319 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 5 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 1/100 dilution
Lane 1 : Human spinal cord tissue lysate - total protein (ab29188)
Lane 2 : Human brain tissue lysate - total protein (ab29466)
Lane 3 : Spinal Cord (Mouse) Tissue Lysate
Lane 4 : Brain (Mouse) Tissue Lysate
Lane 5 : Spinal Cord (Rat) Tissue Lysate
Lane 6 : Brain (Rat) Tissue Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 48 kDa
Exposure time: 1 minute
This antibody was raised against full length native CNPase and is predicted to recognize both isoforms. The predicted molecular weights of isoforms CNPI and CNPII are 45- and 48-kDa respectively. -
Immunocytochemistry - Anti-CNPase antibody [11-5B] (ab6319)This image is courtesy of an anonymous abreview.ab6319 staining CNPase in rat oligodendrocytes by Immunocytochemistry/ Immunofluorescence.
Cells were fixed in paraformaldehyde, blocked using 5% serum for 10 minutes at 25°C, then incubated with ab6319 at a 1/200 dilution for 2 hours at 25°C. The secondary used was a goat anti-mouse Cy3 conjugated polyclonal at a 1/100 dilution. -
Overlay histogram showing SH-SY5Y cells stained with ab6319 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab6319, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNPase antibody [11-5B] (ab6319)
IHC image of CNPase staining in human cerebral cortex formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab6319, 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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Immunocytochemistry - Anti-CNPase antibody [11-5B] (ab6319)This image is courtesy of an anonymous Abreview
ab6319 staining CNPase in the rat oligodendrocytes by ICC/IF (Immunoytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with methanol and blocked with 5% BSA for 1 hour at 37°C. Samples were incubated with primary antibody (1/100 in PBS) for 18 hours at 4°C. An Alexa Fluor® 594-conjugated Goat anti-mouse IgG polyclonal (1:200) was used as the secondary antibody.
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All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 1/750 dilution
All lanes : Spinal Cord homogenate (whole tissue lysate)
Lysates/proteins at 2 µg per lane.
Secondary
All lanes : HRP conjugated sheep anti-mouse IgG
Predicted band size: 48 kDa
Observed band size: 45,47 kDa why is the actual band size different from the predicted?
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ab6319 stained in SKNSH cells. Cells were fixed with 100% methanol (10 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab6319 at 10µg/ml and ab6046 (Rabbit polyclonal to beta Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNPase antibody [11-5B] (ab6319)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom
ab6319 staining CNPase in Dog Cerebellum tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1500 in blocking buffer) for 2 hours at 21°C. A Biotin-conjugated Goat anti-mouset IgG polyclonal (1/200) was used as the secondary antibody.
データシートおよび資料
参考文献 (84)
ab6319 は 84 報の論文で使用されています。
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- Fan H et al. MsrA Suppresses Inflammatory Activation of Microglia and Oxidative Stress to Prevent Demyelination via Inhibition of the NOX2-MAPKs/NF-?B Signaling Pathway. Drug Des Devel Ther 14:1377-1389 (2020). PubMed: 32308370
- Hwang I et al. CIC is a critical regulator of neuronal differentiation. JCI Insight 5:N/A (2020). PubMed: 32229723
- Li S et al. Conversion of Astrocytes and Fibroblasts into Functional Noradrenergic Neurons. Cell Rep 28:682-697.e7 (2019). PubMed: 31315047
- Kano M et al. Tanycyte-Like Cells Derived From Mouse Embryonic Stem Culture Show Hypothalamic Neural Stem/Progenitor Cell Functions. Endocrinology 160:1701-1718 (2019). PubMed: 31135891