概述

• 產(chǎn)品名稱Anti-Neuropilin 1抗體[EPR3113]
     

• 描述

  兔單克隆抗體[EPR3113] to Neuropilin 1

• 經(jīng)測試應(yīng)用WB,IHC-P,IP,Flow Cyt,IHC-Fr,ICC/IF

• 種屬反應(yīng)性

  與反應(yīng): Mouse, Rat, Human, Monkey, Marmoset (common)    
   

• 免疫原

  Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Neuropilin 1 aa 900 to the C-terminus (intracellular).
  (Peptide available as

• 陽性對(duì)照    

• WB: HUVEC and HepG2 cell lysates, human placenta, kidney and heart, mouse heart and kidney and rat heart and kidney tissue lysates. IHC-P: Human liver tissue. ICC/IF: MCF7 and HUVEC cells. Flow Cyt: HepG2 and MCF7 cells.

• 常規(guī)說明

  This product is a recombinant rabbit monoclonal antibody.

  We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

  Produced using Abcam’s RabMAb^? technology. RabMAb^? technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

  Alternative versions available:

  Anti-Neuropilin 1 antibody (Alexa Fluor^? 647) [EPR3113] (ab198323)

  Anti-Neuropilin 1 antibody (Alexa Fluor? 488) [EPR3113] (ab197644)

  Anti-Neuropilin 1 antibody (Phycoerythrin) [EPR3113] (ab209445)

性能

• 形式Liquid

• 存放說明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.

• 存儲(chǔ)溶液pH: 7.20
  Preservative: 0.01% Sodium azide
  Constituents: 59% PBS, 40% Glycerol, 0.05% BSA

• 
  

• 純度Protein A purified

• 克隆單克隆

• 克隆編號(hào)EPR3113

• 同種型IgG

• 研究領(lǐng)域

• Immunology

• Adaptive Immunity

• Regulatory T Cells

• Neuroscience

• Neurology process

• Growth and Development

• Axonal Guidance Proteins

• Cardiovascular

• Angiogenesis

• Growth Factors

• VEGF

• VEGF Receptors

Anti-Neuropilin 1 antibody [EPR3113] 圖像

• 

  Western blot - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)          

     All lanes : Anti-Neuropilin 1 antibody [EPR3113] (ab81321) at 1/10000 dilution (purified)
  
  Lane 1 : Mouse heart tissue lysate
  Lane 2 : Rat heart tissue lysate
  
  Lysates/proteins at 20 μg per lane.
  
  Secondary
  Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
  
  Predicted band size : 103 kDa
  Observed band size : 120 kDa (why is the actual band size different from the predicted?)
     

  Blocking and dilution buffer: 5% NFDM/TBST.

   

• 

  Western blot - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)          

     Anti-Neuropilin 1 antibody [EPR3113] (ab81321) at 1/10000 dilution (purified) + Human heart tissue lysate at 20 μg
  
  Secondary
  Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
  
  Predicted band size : 103 kDa
  Observed band size : 120 kDa (why is the actual band size different from the predicted?)
     

  Blocking and dilution buffer: 5% NFDM/TBST.

   

• 

  Western blot - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)          

     Anti-Neuropilin 1 antibody [EPR3113] (ab81321) at 1/2000 dilution (purified) + Human placenta tissue lysate at 20 μg
  
  Secondary
  Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
  
  Predicted band size : 103 kDa
  Observed band size : 120 kDa (why is the actual band size different from the predicted?)
     

  Blocking and dilution buffer: 5% NFDM/TBST.

   

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)          

     

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Neuropilin 1 with purified ab81321 at 1/400. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

   

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)          

     

  Immunocytochemistry/Immunofluorescence analysis of HUVEC cells labelling Neuropilin 1 with purified ab81321 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^? 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^? 594-conjugated goat anti-mouse IgG (1/500) were also used.

  Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor^? 594-conjugated goat anti-mouse IgG (1/500).

  Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^? 488-conjugated goat anti-rabbit IgG (1/500).

   

• 

  Flow Cytometry - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)          

     

  Flow Cytometry analysis of MCF7 cells labelling Neuropilin 1 with purified ab81321 at 1/70 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

   

• 

  Immunohistochemistry (Frozen sections) - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)      Image courtesy of an anonymous Abreview.    

     

  Immunohistochemical analysis of frozen human kidney tissue sections labelling Neurophilin 1 with ab81321 at a concentration of 1/100 for 18 hours at 4°C. The antibody was then blocked with a serum free protein block for 1 hour at 21°C. The secondary antibody used was a donkey anti-rabbit antibody conjugated to an Alexa488^? dye inclubated at 1/400.

     

  See Abreview

• 

  Western blot - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)          

     All lanes : Anti-Neuropilin 1 antibody [EPR3113] (ab81321) at 1/1000 dilution (unpurified)
  
  Lane 1 : Human placenta lysate
  Lane 2 : HUVEC cell lysate
  Lane 3 : HepG2 cell lysate
  Lane 4 : Mouse heart tissue lysate
  Lane 5 : Mouse kidney tissue lysate
  Lane 6 : Rat heart tissue lysate
  Lane 7 : Rat kidney tissue lysate
  
  Lysates/proteins at 10 μg per lane.
  
  Secondary
  HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
  
  Predicted band size : 103 kDa
  Observed band size : 120 kDa (why is the actual band size different from the predicted?)
   

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)      This image is courtesy of an Abreview submitted by Manoj Kumar Valluru    

     

  Unpurified ab81321 staining Neuropilin 1 in mouse brain tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/100 in PBS + 2% blocking serum) for 16 hours at 4°C. A biotin-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

     

  See Abreview

• 

  Immunohistochemistry (Frozen sections) - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)      Image courtesy of an anonymous Abreview.    

     

  Unpurified ab81321 staining Neuropilin 1 in rat brain tissue sections by Immunohistochemistry (frozen sections). Tissue was fixed with paraformaldehyde and then blocked with 10% serum for 1 hour at 27°C followed by incubation with the primary antibody, undiluted, for 14 hours at 4°C. An undiluted Cy3^? conjugated donkey anti-rabbit was used as the secondary antibody.

     

  See Abreview

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)          

     

  ICC/IF image of unpurified ab81321 stained MCF7 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 (unpurified ab81321, 5μg/ml) overnight at +4°C. The secondary antibody (green) was DyLight^? 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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.

   

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)      This image is courtesy of an anonymous Abreview    

     

  Unpurified ab81321 staining Neuropilin 1 in the COS1 fibroblast-like cell line derived from monkey kidney tissue by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 0.1% and blocked with 10% serum for 60 minutes at 24°C. Samples were incubated with primary antibody (1/200) for 16 hours at 4°C. An Alexa Fluor^? 488-conjugated goat anti-rabbit monoclonal(1/500) was used as the secondary antibody.

     

  See Abreview

• 

  Flow Cytometry - Anti-Neuropilin 1 antibody [EPR3113] (ab81321)          

     

  Overlay histogram showing HepG2 cells stained with unpurified ab81321 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (unpurified ab81321, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^? 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HepG2 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.