Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker

• 产品名称

  Anti-160 kD Neurofilament Medium抗体[NF-09] - Neuronal Marker
  参阅全部 160 kD Neurofilament Medium 一抗

• 描述

  小鼠单克隆抗体[NF-09] to 160 kD Neurofilament Medium - Neuronal Marker

• 宿主

  Mouse

• 特异性

  This antibody reacts with both phosphorylated and non-phosphorylated forms of medium neurofilament protein (160 kDa) of various species.

• 经测试应用

  适用于: WB, ELISA, ICC/IF, IHC-Fr, IHC-P, IHC (PFA fixed)more details

• 种属反应性

  与反应: Mouse, Rat, Cow, Cat, Human, Pig
  预测可用于: a wide range of other species, Mammals

• 免疫原

  Pellet of pig brain cold stable proteins after depolymerization of microtubules.

• 阳性对照

• WB: HEK-293 and A549 whole cell lysates ICC/IF: Dental pulp stem, Neuro2A, PC12 and human nerve cells

• 常规说明

  
  

  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: 0.097% Sodium azide
  Constituent: PBS

• 纯度

  Protein A purified

• 克隆

  单克隆

• 克隆编号

  NF-09

• 同种型

  IgG2a

• 轻链类型

  unknown

The Abpromise guarantee

Abpromise™承诺保证使用ab7794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: 281347 Cow

• Entrez Gene: 4741 Human

• Entrez Gene: 18040 Mouse

• Entrez Gene: 24588 Rat

• Omim: 162250 Human

• SwissProt: O77788 Cow

• SwissProt: P07197 Human

• SwissProt: P08553 Mouse

• SwissProt: P12839 Rat

• Unigene: 458657 Human

• Unigene: 730072 Human

• Unigene: 242832 Mouse

• Unigene: 390700 Mouse

• Unigene: 10971 Rat

别名

• 150kDa medium antibody

• 160 kDa neurofilament protein antibody

• NEF3 antibody

• NEFM antibody

• Neurofilament 3 antibody

• Neurofilament medium polypeptide antibody

• Neurofilament triplet M protein antibody

• NF-M antibody

• NF160 antibody

• NFM antibody

• NFM_HUMAN antibody

• 

  Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)

  All lanes : Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794) at 1/1000 dilution
  
  Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
  Lane 2 : NEFM knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
  Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
  
  Observed band size: 150 kDawhy is the actual band size different from the predicted?
  
  
  
  

  Lanes 1-3: Merged signal (red and green). Green - ab7794 observed at 150 kDa. Red - loading control ab181602 observed at 36 kDa.

   ab7794 Anti-160 kD Neurofilament Medium antibody [NF-09]  was shown to specifically react with 160 kD Neurofilament Medium in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266741 (knockout cell lysate ab257103) was used. Wild-type and 160 kD Neurofilament Medium knockout samples were subjected to SDS-PAGE. ab7794 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)Image from Ferro F et al., PLoS One. 2012;7(7):e41774. Epub 2012 Jul 23. Fig 3.; doi:10.1371/journal.pone.0041774; July 23, 2012, PLoS ONE 7(7): e41774.

  Immunofluorescence analysis of 1 month neuronal-differentiated dental pulp stem cells, staining 160 kD Neurofilament Medium with ab7794.
  
  Cells were fixed with paraformaldehyde and incubated with primary antibody (1/600). A FITC-conjugated anti-mouse IgG (1/375) was used as the secondary antibody.

• 

  Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)

  All lanes : Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794) at 1/1000 dilution
  
  Lane 1 : Wild-type HEK-293 whole cell lysate
  Lane 2 : NEFM knockout HEK-293 whole cell lysate
  Lane 3 : A549 whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Observed band size: 150 kDawhy is the actual band size different from the predicted?
  
  
  
  

  Lanes 1 - 4: Merged signal (red and green). Green - ab7794 observed at 150 kDa. Red - loading control, ab52866, observed at 50 kDa.

  ab7794 was shown to specifically react with NEFM (Neurofilament) in wild-type HEK-293 cells as signal was lost in NEFM knockout cells. Wild-type and NEFM knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab7794 and ab52866 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1/1000 dilution 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.

   

• 

  Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)

  Western blotting analysis of neurofilament medium protein in porcine brain lysate (reducing conditions) by mouse monoclonal NF-09.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)

  ab7794 staining Neurofilament medium protein in mouse Neuro2A cells by ICC/IF.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)

  ICC/IF image of ab7794 stained PC12 cells. The cells were 100% methanol fixed (5 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 (ab7794, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-mouse 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 1.43æM.