Anti-CNPase antibody [11-5B]

• 产品名称

  Anti-CNPase抗体[11-5B]
  参阅全部 CNPase 一抗

• 描述

  小鼠单克隆抗体[11-5B] to CNPase

• 宿主

  Mouse

• 经测试应用

  适用于: IHC-P, WB, ICC/IFmore details

• 种属反应性

  与反应: Mouse, Rat, Human
  预测可用于: Sheep, Rabbit, Cow, Dog, Pig, Rhesus monkey不与反应: Chicken, Guinea pig

• 免疫原

  Full length native protein (purified) corresponding to Human CNPase.

• 阳性对照

• 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: Primary hippocampal rat neurons/glia, DIV14. cells

• 常规说明

  
  

  This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

• 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 or -80°C. Avoid freeze / thaw cycle.

• 存储溶液

  pH: 7.40
  Preservative: 0.02% Sodium azide
  Constituents: PBS, 6.97% L-Arginine

• 纯度

  Affinity purified

• 克隆

  单克隆

• 克隆编号

  11-5B

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab6319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。

数据库链接

• 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

• SwissProt: P06623 Cow

• SwissProt: P09543 Human

• SwissProt: P16330 Mouse

• SwissProt: P18104 Pig

• SwissProt: P13233 Rat

• SwissProt: P56283 Sheep

• Unigene: 273621 Human

• Unigene: 15711 Mouse

• Unigene: 31762 Rat

别名

• 2'' antibody

• 2'3' cyclic nucleotide 3' phosphodiesterase antibody

• 3''-cyclic-nucleotide 3''-phosphodiesterase antibody

• CN37_HUMAN antibody

• CNP 1 antibody

• CNP antibody

• CNP1 antibody

• CNPase antibody

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CNPase antibody [11-5B] (ab6319)

  ab6319 staining CNPase in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab6319 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 4% paraformaldehyde (10 min).

  Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• 

  Western blot - Anti-CNPase antibody [11-5B] (ab6319)

  All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 5 µg/ml
  
  Lane 1 : Wild-type HAP1 whole cell lysate at 40 µg
  Lane 2 : CNPase knockout HAP1 whole cell lysate at 40 µg
  Lane 3 : Human Brain whole cell lysate at 20 µg
  
  Predicted band size: 48 kDa
  
  
  
  

  Lanes 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.

• 

  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.

  See Abreview

• 

  Western blot - Anti-CNPase antibody [11-5B] (ab6319)

  All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 5 µg/ml
  
  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 kDa
  
  
  
  

  Lanes 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.

• 

  Western blot - Anti-CNPase antibody [11-5B] (ab6319)

  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/ Immunofluorescence - 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.

  See Abreview

• 

  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.

• 

  Immunocytochemistry/ Immunofluorescence - 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.

  See Abreview

• 

  Western blot - Anti-CNPase antibody [11-5B] (ab6319)This image is courtesy of an anonymous Abreview

  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 kDawhy is the actual band size different from the predicted?
  
  

  See Abreview

• 

  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.

  See Abreview