Overview

  • Product name
    Anti-Nestin antibody [10C2]
    See all Nestin primary antibodies
  • Description
    Mouse monoclonal [10C2] to Nestin
  • Host species
    Mouse
  • Specificity
    Reacts exclusively with human nestin protein.
  • Tested applications
    Suitable for: ICC/IF, IHC-Fr, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    A recombinant 150 amino acid fragment from human Nestin conjugated to glutathione S transferase.

  • Positive control
    • WB: U251 cells. ICC/IF: U251 cells. Human brain tissue.
  • General notes

    Monoclonal antibody 10C2 was derived by fusion of P3x63Ag8.653 myeloma cells with spleen cells from a BALB/c mouse immunized with a 150-amino-acid fragment from the cloned human nestin, conjugated to glutathione S-transferase.

Properties

Applications

Our Abpromise guarantee covers the use of ab22035 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 1/100 - 1/200. PubMed: 19147502

1/1000.

IHC-Fr 1/100 - 1/200.
IHC-P Use at an assay dependent concentration.
WB 1/1000 - 1/5000. Predicted molecular weight: 176 kDa.

1/1000 - 1/5000 using chemiluminescent detection. Predicted molecular weight: 176 kDa.

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

  • ab22035 staining human brain tissue cells by ICC/IF.  Cells were PFA fixed and permeabilized in Triton X-100 prior to blocking in 1% BSA for 30 minutes at 21°C.  The primary antibody was diluted 1/100 and incubated with the samples for 1 hour at 21°C.  A FITC conjugated goat anti-mouse IgG antibody, diluted 1/250, was used as the secondary.

    See Abreview

  • ICC/IF of U251 cells staining intracellular Nestin filaments with ab22035

  • Lane 1 : Anti-Nestin antibody [10C2] (ab22035) at 1/1000 dilution
    Lane 2 : Anti-Nestin antibody [10C2] (ab22035)

    Lane 1 : U251 cells at 10 µl
    Lane 2 : U251 cells at 20 µl

    Predicted band size: 176 kDa


    Exposure time: 60 seconds

References

This product has been referenced in:
  • Wang G  et al. Human amniotic epithelial cells regulate osteoblast differentiation through the secretion of TGFß1 and microRNA-34a-5p. Int J Mol Med 41:791-799 (2018). IHC-P ; Human . Read more (PubMed: 29207015) »
  • Liu J  et al. Nestin-expressing cell types in the temporal lobe and hippocampus: Morphology, differentiation, and proliferative capacity. Glia 66:62-77 (2018). IHC-P ; Human . Read more (PubMed: 28925561) »
See all 50 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (First trimester human placenta)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate
Permeabilization
No
Specification
First trimester human placenta
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 03 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (bone marrow-mesenchymal stem cell)
Permeabilization
Yes - saponin 0,1%
Specification
bone marrow-mesenchymal stem cell
Blocking step
Protein block serum-free ready to use (DAKO) + saponin 0,1% as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 23 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human iPSC derived neural stem cell)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Human iPSC derived neural stem cell
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 13 2015

Answer

Thank you for contacting us.

1. Anti-Nestin antibody [10C2] (ab22035) and Anti-HIF1 alpha antibody [H1alpha67] - BSA and Azide free (ab80544):

Volume: 100 µg at 1 mg/ml

We recommend to start with a concentration of 5 ug/ml. This will need further optimisation depending on samples and protocol.

2. Anti-c-Myc antibody [Y69] (ab32072)

Volume: 100 ul with a concentration from 0.1120 mg/ml for lot GR85451-5 (please refer to this lot when ordering).

We recommend to use this antibody in a dilution of 1/50 in IHC-P. As said previously this recommendation will need optimisation for every experiment by user.

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 very much for your interest in anti-Nestin antibodies.

To our knowledge our anti-Nestin products have not been tested in bovine. Therefore, I can offer a discount off a future purchase if you buy an anti-Nestin antibody now, test it in bovine for ICC and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free PRIMARY ANTIBODY.

While we do have 18 products against this target, I would suggest the following. These are either well characterized in the application or have been used in a number of species. I ordered them in what I feel is the most appropriate first but I can offer this discount for any or all of these.

ab6142: https://www.abcam.com/Nestin-antibody-2Q178-Neural-Stem-Cell-Marker-ab6142.html
ab27952: https://www.abcam.com/Nestin-antibody-Azide-free-ab27952.html
ab22035: https://www.abcam.com/Nestin-antibody-10C2-ab22035.html
ab81755: https://www.abcam.com/Nestin-antibody-ab81755.html

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test one or more of these antibodies in ICC on bovine. I will then send a discount code. This code must be issued before purchase so please wait for my reply before ordering.

2. Purchase either by phone, fax, or online (www.abcam.com).

3. Test it in bovine.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if the antibody turns out to be unsuitable for bovine, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you very much for your email. I apologize for the delay in our answer.

I can confirm thatthis antibody can be used with HRP conjugated secondary antibody without any problems.

The note on the datasheet is in reference on how it has been tested: the ab22035 has been tested on paraffin embedded tissue slides witha secondary antibody conjugated to FITC or Rhodamine. Inthat case, adilution of the ab22035 at1/100 to 1/200 gave good signal.

I would expect therefore, that using this antibody at 1/100 to 1/200 will also give good results using a secondary antibody coupled to HRP or the EXPOSE kit.

I hope this information is helpful. Please do not hesitate to contact us againshould you need any further information.

Read More
Application
Flow Cytometry
Sample
Human Cell (Differentiation Human ES cell)
Specification
Differentiation Human ES cell
Preparation
Cell harvesting/tissue preparation method: Cell dissociation buffer
Sample buffer: FACS buffer (0.2%BSA, 0.1%sodium azide in PBS)
Fixation
Formaldehyde
Permeabilization
Yes - 70% methanol (ice cold)
Gating Strategy
FSC vs SSC on negative control sample

Abcam user community

Verified customer

Submitted Jan 09 2012

Answer

Thank you for your patience. I have attached the test results from the lab. We performed an immunoblotting experiment on a cell extract of U251 cells. The attached Western blot image clearly indicates a Nestin band with a MW of approximately 250 kDa. The test was a result of a 1:1000 antibody dilution prepared from a purified stock solution of 1mg/ml. The protein might be run higher than theoretically predicted due to the large number of phosphorylation sites (about 30). This however would not explain why you see a band at only 90 kDa. Please let me know if you have tried anything else in the meantime and receivd different results. If not and the antibody was ordered within the last 6 month, we can discuss a replacement, credit or refund. I hope this information is nevertheless helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (brain tumor cell line)
Specification
brain tumor cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.05% Triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 29 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human ES cell derived neurons)
Specification
human ES cell derived neurons
Fixative
Paraformaldehyde
Permeabilization
Yes - See Blocking buffer
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jun 21 2011

1-10 of 13 Abreviews or Q&A

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