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 22 Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews

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

Abcam user community

Verified customer

Submitted Dec 18 2013

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

Ms. Erica Sloma

Verified customer

Submitted Aug 21 2014

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

Abcam user community

Verified customer

Submitted Nov 22 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Nov 22 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Hongwei Shao

Verified customer

Submitted Nov 09 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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