PDF《Product datasheet》

Product name Anti-Nestin antibody [SP103] Description Rabbit monoclonal [SP103] to Nestin Host species Rabbit Tested applications Suitable for: ICC/IF, Flow Cyt, WB, IHC-P Species reactivity Reacts with: Human Predicted to work with: Dog Immunogen Synthetic peptide within Human Nestin aa

1600 to the C-terminus (C terminal).

The exact sequence is proprietary. Database link: P48681 Positive control IHC-P: Human kidney, ovary, cervix, breast, colon, endometrium, cerebellum, lung, prostate, skin and human melanoma tissue. WB: HeLa cell lysate. Flow Cyt: SH-SY5Ycells. ICC/IF: SKNSH cells. General notes This product is a recombinant rabbit monoclonal antibody. Form Liquid Storage instructions Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. Storage buffer pH: 7.60 Preservative: 0.1% Sodium azide Constituents: PBS, 1% BSA Purity Protein A/G purified Purification notes Purified from TCS by protein A/G. Clonality Monoclonal Clone number SP103 Isotype IgG Product datasheet Product datasheet Anti-Nestin antibody [SP103] ab105389 Anti-Nestin antibody [SP103] ab105389 Recombinant

5 Abreviews

22 References

16 Images Overview Properties

1 Function Required for brain and eye development. Promotes the disassembly of phosphorylated vimentin intermediate filaments (IF) during mitosis and may play a role in the trafficking and distribution of IF proteins and other cellular factors to daughter cells during progenitor cell division. Required for survival, renewal and mitogen-stimulated proliferation of neural progenitor cells. Tissue specificity CNS stem cells. Sequence similarities Belongs to the intermediate filament family. Developmental stage Upon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments. Post-translational modifications Constitutively phosphorylated. This increases during mitosis when the cytoplasmic intermediate filament network is reorganized. Western blot - Anti-Nestin antibody [SP103] (ab105389) All lanes : Anti-Nestin antibody [SP103] (ab105389) at 1/100 dilution Lane

1 : Wild-type HAP1 whole cell lysate Lane

2 : NES knockout HAP1 whole cell lysate Lysates/proteins at

20 ?g per lane. Predicted band size:

177 kDa Lanes

1 - 2: Merged signal (red and green). Green - ab105389 observed at

300 kDa. Red - loading control, ab18058, observed at Applications Our Abpromise guarantee covers the use of ab105389 in the following tested applications. The application notes include recommended starting dilutions;

optimal dilutions/concentrations should be determined by the end user. Application Abreviews Notes ICC/IF 1/200. Flow Cyt 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. WB 1/100. Predicted molecular weight:

177 kDa. IHC-P 1/100. Antigen retrieval: Boil tissue section in 10mM citrate buffer, pH 6.0 for

10 min followed by cooling at room temperature for

20 min. Target Images

2 117 kDa. ab105389 was shown to specifically react with NES in wild-type HAP1 cells as signal was lost in NES knockout cells. Wild-type and NES knockout samples were subjected to SDS-PAGE. Ab105389 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/100 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&

L (IRDye 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&

L (IRDye 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for

1 hour at room temperature before imaging. Immunocytochemistry/ Immunofluorescence - Anti- Nestin antibody [SP103] (ab105389) ICC/IF image of ab105389 stained SKNSH cells. The cells were 4% formaldehyde 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 (ab105389, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight?