Overview

  • Product name
    Anti-Nestin antibody [SP103]
    See all Nestin primary antibodies
  • Description
    Rabbit monoclonal [SP103] to Nestin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, WB, IHC-Pmore details
  • 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-SY5Y cells. ICC/IF: SKNSH cells.
  • General notes

     

     

    This product is a recombinant rabbit monoclonal antibody.

Properties

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

  • 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.
  • Information by UniProt
  • Database links
  • Alternative names
    • ESTM 46 antibody
    • FLJ 21841 antibody
    • FLJ21841 antibody
    • Intermediate filament protein antibody
    • Nbla00170 antibody
    • nes antibody
    • NEST_HUMAN antibody
    • Nestin antibody
    see all

Images

  • 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 µg 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 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.

  • 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® 488 goat anti-rabbit IgG (H+L) 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.
  • BTIC markers in our CNS-PNET model tumors

    Formalin-fixed, paraffin-embedded mouse primitive neural ectodermal tumor tissue was stained for Nestin using ab105389 at 1/30 dilution in immunohistochemical analysis.

    Insert x20 magnification.

    TU-tumor; P-parenchyma; V-ventricle; CP-choroid plexus

    (From Figure 3C of Malchenko et al)

  • Formalin-fixed, paraffin-embedded human endometrium tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.

  • Anti-Nestin antibody [SP103] (ab105389) at 1/100 dilution + HeLa cell lysate

    Predicted band size: 177 kDa

  • Overlay histogram showing SH-SY5Y cells stained with ab105389 (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 (ab105389, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Formalin-fixed, paraffin-embedded human colon tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human prostate tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human lung tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human breast tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human skin tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human cerebellum tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human cervix tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human kidney tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human ovary tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.

  • ab105389 staining Nestin in human melanoma tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using Tris EDTA pH 9.0. Samples were then permeabilized using 0.25% Triton X-100, blocked, then incubated with ab105389 at a 1/100 dilution for 18 hours at 4°C. The secondary used was an Alexa-Fluor 594 (red) conjugated donkey anti-rabbit polyclonal used at a 1/100 dilution. (Green) tumor cells, (blue) DAPI.

    See Abreview

References

This product has been referenced in:
  • Mesci P  et al. Blocking Zika virus vertical transmission. Sci Rep 8:1218 (2018). Read more (PubMed: 29352135) »
  • Colini Baldeschi A  et al. Atrial Natriuretic Peptide Acts as a Neuroprotective Agent in in Vitro Models of Parkinson's Disease via Up-regulation of the Wnt/ß-Catenin Pathway. Front Aging Neurosci 10:20 (2018). WB, ICC/IF . Read more (PubMed: 29449807) »
See all 25 Publications for this product

Customer reviews and Q&As

1-3 of 3 Q&A

Answer

Thank you for contacting us. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab105389. I would also appreciate if you can confirm some further details:

1) How long did you permeabilize with Triton?

2) Overnight incubation was used in the image on our datasheet. This is usually the best way to get a strong signal in your cells.

Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

Read More

Answer

Thank you for contacting us.

Unfortunately, alignments of the immunogen with mouse show very poor homology. As such, we cannot guarantee or recommend this product for use in this species. We do offer a number of other anti-Nestin antibodies of which nine are tested and guaranteed in mouse samples. In general, I would recommend Anti-Nestin antibody [2Q178] - Neural Stem Cell Marker (ab6142) as this product is extensively tested, published with and reviewed. However, I would be happy to help find the right product for you based on your needs.

More information about ab6142 may be found at this location: https://www.abcam.com/nestin-antibody-2q178-neural-stem-cell-marker-ab6142.html

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Answer

Thank you for contacting us.

The concentration of lot in question is XXXug/ml (or XX mg/ml).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up