Overview

  • Product name
    Anti-Nestin antibody [196908]
    See all Nestin primary antibodies
  • Description
    Mouse monoclonal [196908] to Nestin
  • Host species
    Mouse
  • Specificity
    Based on the result of immunocytochemistry on mouse and rat neural stem cells, this antibody has no reactivity with rodent Nestin. The clone number has been updated from (3k1) to (196908) both clone numbers name the same antibody clone.
  • Tested applications
    Suitable for: IHC-Fr, IHC - Wholemount, IHC-FoFr, ICC, Flow Cyt, ELISA, WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    Nestin-transfected NS0 cells transfected with a human Nestin fragment aa residues 618-1618.

  • Positive control
    • Human neural progenitors and A172 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab6320 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
IHC-Fr Use a concentration of 0.1 µg/ml. When using aldehyde-based fixative, use at a concentration of 5 µg/ml (see Abreview).
IHC - Wholemount Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration.
ICC Use a concentration of 5 - 10 µg/ml. Tested in human neural progenitors and A172 cells. Cells were fixed with 4% paraformaldehyde and 0.15% picric acid in PBS at RT for 20 min.
Flow Cyt Use a concentration of 25 µg/ml.

Fix and permeabilize cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ELISA Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

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

  • Immunocytochemical analysis of human fetal neural progenitor cells, labeling Nestin with ab6320 (10µg/ml). Immunostaining was for 3 hours at room temperature. Counterstaining with DAPI (blue).

  • Tumorigenicity of human glioma-derived cells. Representative immunohistochemical analysis of brain tumors generated after orthotropic injection of isolated human glioblastoma cells. Immunofluorescence of mouse brains cryosections labeled with anti-human nestin antibody. A, diffuse infiltration of the injected and contralateral hemisphere (GBM 1). B, cells infiltrating the striatum ipsilateral and contralateral to the side of injection (GBM 2). Scale bar, 100 µm. GBM xenografts recapitulate the morphology of corresponding tumors in human cancer patients.
    For xenograft tumor analysis, mice were sacrificed, and cryopreserved brain sections were cut using a 10-µm cryostat. Staining with hematoxylin-eosin identified sections bearing tumors. Briefly, brain sections were mounted on slides and stained with hematoxylin for 40 seconds and then counterstained with alcoholic eosin for 30 seconds. Cryosections containing tumors were permeabilized in PBS containing 0.5% Triton X-100 and
  • ab6320 staining Nestin in Human glioblastoma in mouse brain by Immunohistochemistry - Wholemount.

    Samples were incubated with primary antibody (1/1000 in diluent) for 16 hours at 4°C. An HRP-conjugated goat anti-mouse polyclonal IgG was used as the secondary antibody.

References

This product has been referenced in:
See all 20 Publications for this product

Customer reviews and Q&As

1-10 of 14 Abreviews or Q&A

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Antigen retrieval step
None
Sample
Human Tissue sections (human tumor bearing mouse brain)
Specification
human tumor bearing mouse brain
Blocking step
bsa 1% + Goat serum 5% as blocking agent for 2 hour(s) and 0 minute(s) · Temperature: 20°C
Fixative
Paraformaldehyde

Mrs. Francoise Geffroy

Verified customer

Submitted May 02 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Sample
Human Cell (hair follicle culture)
Specification
hair follicle culture
Permeabilization
Yes - methanol
Fixative
Paraformaldehyde

Mr. Heiko Locher

Verified customer

Submitted Sep 23 2013

Application
Immunocytochemistry
Sample
Mouse Cell (adipose derived stem cell)
Specification
adipose derived stem cell
Fixative
Formaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 03 2013

Answer

Thank you very much for your call yesterday, and I apologize for the variation of results using this antibody.

As we discussed, I'm sending two free of charge vials to Cedarlane, who will then forward the replacement vials on to you.

Please keep me updated about the results using the new vials, and let me know if you have any questions or need anything else. Have a nice day!

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Primary glioblastoma cell lines)
Loading amount
25 µg
Specification
Primary glioblastoma cell lines
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Aug 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
IHC - Wholemount
Sample
Human Tissue (Human glioblastoma in mouse brain)
Specification
Human glioblastoma in mouse brain

Abcam user community

Verified customer

Submitted Jul 26 2012

Answer

Thank you for your reply with this information.


Regarding your protocol - one thing that really stood out is the use of Triton as a permeabilizing agent. This detergent will also partially dissolve the nuclear membrane and will disrupt proteins when used at higher concentrations for longer periods of time. We typically recommend using it at 0.1-0.2% for 10 min only. Since both of these proteins are cytomplasmic, permeabilization with tween 20 is sufficient (I recommend 0.2 % for 10 - 15 minutes). This should decrease the amount of background and non-specific stain you are observing.


Additionally, I would recommend optimizing the conditions with each antibody individually rather than in sequential staining.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or concerns.

Read More

Answer

Thank you for contacting Abcam.


I am sorry that you have been experiencing difficulties with these antibodies in ICC. I have reviewed the images provided, but I was wondering if you would also provide a detailed summary of your protocol and sample preparation. This will help me to evaluate if there are any suggestions I can make to improve your results or if the antibodies are faulty.


I look forward to your reply so that I may assist you further. Please do not hesitate to contact me with any additional questions or concerns.

Read More

Question
Answer

Thanks for your call today and for letting us know about the problem with ab6320. As we discussed, I am sending a free of charge vial of ab22035 on the order ***, which I expect to arrive tomorrow. Please keep me updated of the results with this replacement antibody. If you have any images to send, you can just reply to this email and it will go straight to me. This will help us determine how to fix the issue with ab6320. Please let me know if you have any questions or if there is anything else that we can do for you.

Read More

Answer

Thank you for your enquiry. Ab27952 is a rabbit polyclonal antibody to nestin, and ab6320 is a mouse monoclonal antibody. The immunogen used to generate ab27952 was a KLH conjugated peptide to the human nestin tail. The immunogen used to generate ab6320 were nestin-transfected NS0 cells transfected with a human Nestin fragment aa residues 618-1618. Ab27952 detects human and mouse nestin while ab6320 detects human nestin and has no reactivity with rodent nestin. Please contact us again if you have any additional questions.

Read More

1-10 of 14 Abreviews or Q&A

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