Product nameAnti-Nestin antibody 
See all Nestin primary antibodies
DescriptionMouse monoclonal  to Nestin
SpecificityBased 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 applicationsSuitable for: IHC-Fr, IHC - Wholemount, IHC-FoFr, ICC, Flow Cyt, ELISA, WB, IP, ICC/IFmore details
Species reactivityReacts with: Human
Does not react with: Mouse, Rat
Nestin-transfected NS0 cells transfected with a human Nestin fragment aa residues 618-1618.
- Human neural progenitors and A172 cells.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferPreservative: 0.09% Sodium Azide
Constituents: 50% Glycerol, PBS, pH 7.2
Concentration information loading...
PurityProtein G purified
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.
|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.
|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.|
FunctionRequired 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 specificityCNS stem cells.
Sequence similaritiesBelongs to the intermediate filament family.
Developmental stageUpon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments.
modificationsConstitutively phosphorylated. This increases during mitosis when the cytoplasmic intermediate filament network is reorganized.
- Information by UniProt
- ESTM 46 antibody
- FLJ 21841 antibody
- FLJ21841 antibody
All lanes : Anti-Nestin antibody  (ab6320) at 1 µg
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : NES knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Lanes 1 - 2: Merged signal (red and green). Green - ab6320 observed at 300 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab6320 was shown to recognize NES in wild-type HAP1 cells as signal was lost at the expected MW in NES knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NES knockout samples were subjected to SDS-PAGE. Ab6320 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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.
This product has been referenced in:
- Russell OM et al. Preferential amplification of a human mitochondrial DNA deletion in vitro and in vivo. Sci Rep 8:1799 (2018). Read more (PubMed: 29379065) »
- Zhao C et al. ETV2 mediates endothelial transdifferentiation of glioblastoma. Signal Transduct Target Ther 3:4 (2018). Read more (PubMed: 29527330) »