Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Nestin antibody [SP103] (ab105389)

Reviews (5)Q&A (3)References (22)

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

    • Western blot - Anti-Nestin antibody [SP103] (ab105389)
      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 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Nestin antibody [SP103] (ab105389)
      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® 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.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)Malchenko, S. et al PLoS One. 2017 Mar 1;12(3):e0173106. doi: 10.1371/journal.pone.0173106. eCollection 2017

      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)

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

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

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

      Predicted band size: 177 kDa

    • Flow Cytometry - Anti-Nestin antibody [SP103] (ab105389)
      Flow Cytometry - Anti-Nestin antibody [SP103] (ab105389)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)This image is courtesy of Hongwei Shao by Abreview.
      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 22 Publications for this product

    Publishing research using ab105389? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-8 of 8 Abreviews or Q&A

    Western blot abreview for Anti-Nestin antibody [SP103]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    5 µg
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Human Cell lysate - other (undifferentiated human iPS cells)
    Specification
    undifferentiated human iPS cells
    Blocking step
    5% milk in TBST as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 18 2013

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nestin antibody [SP103]

     Inconclusive
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Dako protein block as blocking agent for 10 minute(s) · Concentration: .25% · Temperature: 22°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate
    Sample
    Dog Tissue sections (Fetal tissue)
    Specification
    Fetal tissue
    Permeabilization
    No
    Fixative
    Formaldehyde
    Read More

    Ms. Erica Sloma

    Verified customer

    Submitted Aug 21 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Nestin antibody [SP103]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    1% serum, 0.1% triton, 0.1% BSA in PBS as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
    Sample
    Human Cell (neural progenitor cell derived from human iPSCs)
    Specification
    neural progenitor cell derived from human iPSCs
    Permeabilization
    Yes - (see blocking buffer)
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 22 2013

    Question


    Inquiry: I ordered the RabMAb Anti-nestin antibody and I tested it using conditions recommended in the spec sheet for ICC/IF (1:200). I used this antibody at the same time as some other nestin antibodies from other companies, so I know the cells (human glioblastoma stem cells) express a lot of nestin. And I was also testing the RabMab anti-GFAP antibody (ab68428) at the same time, and this antibody worked really well. I have since repeated the experiment using different antibody concentrations and still have negative results (I get no signal). I am using the Alexafluor goat anti- rabbit 488 secondary antibody and again I was testing other rabbit antibodies at the same time and it worked at the concentration I used (1:250). My protocol is: 1. Fix in 8% PFA added directly to cells in culture media in a 1:1 ratio so it's diluted to 4% 2. wash 3. Permeabilize with 0.1% Triton-x-100 4. wash 5. Block in 10% goat serum 6. Add primary antibody in blocking solution for 1 hr at RT (I've gone as low as 1:100) 7. wash 8. Add secondary antibody in blocking solution for 1 hr at RT 9. wash 10. Add Hoescht 33342 I tried to be as detailed as possible, but please let me know if you need anymore info about my experiment. Thanks for your help.

    Read More
    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

    Question

    I have a question regarding one of you anti-Nestin antibodies ( ab105389 ).  On the website it lists this antibody as reacting with human, but doesn't provide any informatin regarding mouse.  I looked at a number of other Nestin antibodies and some react with both mouse and human, and some are specific to one or the other (for example a mouse ab that lists as "doesn't react with human").  I was wondering if based on the antigen used for this antibody if it if would or wouldn't react with mouse.

    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

    Question

    Please see customers request below:

    Can you please kindly provide me with the information regarding the protein concentration of the following antibody:

    Anti-nestin antibody, cat number = ab105389, Lot = GR36801-1

    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!
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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nestin antibody [SP103]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (kidney)
    Specification
    kidney
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA pH9, 20 min, 100C
    Permeabilization
    No
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 22 2011

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nestin antibody [SP103]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Melanoma)
    Specification
    Melanoma
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris EDTA pH9.0
    Permeabilization
    Yes - 0.25% Triton X-100
    Blocking step
    DakoCytomation X0909 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
    Read More

    Hongwei Shao

    Verified customer

    Submitted Nov 09 2011

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