Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Drosophila, Xenograft

Applications

Validated:

Intracellular Staining by Flow Cytometry, Immunocytochemistry, CyTOF-reported

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunofluorescence, Immunocytochemistry, Bioassay, Cell Culture

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 196908
View all Nestin Primary Antibodies »

Product Specifications

Immunogen

NS0 mouse myeloma cell line transfected with human Nestin

Specificity

Detects human Nestin.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human Nestin Antibody

Nestin antibody in Human Neural Progenitor Cells by Immunocytochemistry (ICC).

Nestin in Human Neural Progenitor Cells.

Nestin was detected in immersion fixed human fetal neural progenitor cells using 10 µg/mL Human Nestin Monoclonal Antibody (Catalog # MAB1259) for 3 hours at room temperature. Cells were stained (green) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Nestin in A172 cells by Flow Cytometry.

A172 cells were stained with Mouse Anti-Human Nestin Monoclonal Antibody (Catalog # MAB1259, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Detection of Mouse Nestin by Western Blot

Detection of Mouse Nestin by Western Blot

Cardiolipin is externalized to the mitochondrial surface in alpha -syn mutant hNs and transgenic mice in response to alpha -syn accumulation. a Western analysis of lysates from corrected and A53T cells at DIV 14 and DIV 60 labeled for total alpha -syn, PS129, TH (DA neuronal marker), nestin (NPC marker) or GAPDH show that levels of PS129-modified alpha -syn are elevated in A53T hNs at DIV 14 and remain elevated at DIV 60 relative to corrected cells. b, c Mitochondria were purified from A53T cells and genetically corrected cells at both DIV 14 and DIV 60. Lysates were then separated based on TX-100 solubility (soluble) or urea solubility (insoluble). Western analysis of A53T cells shows elevated soluble alpha -syn at the mitochondria relative to corrected cells at DIV 14 and DIV 60 (denoted by ** and * respectively). Quantification of soluble and insoluble alpha -syn levels in mitochondrial fractions normalized to Coomassie (c). Data represent mean ± s.e.m. *P < 0.05, **P < 0.01 by ANOVA followed by Tukey’s post hoc test, n = 4. d, e Micrographs of corrected and A53T-mutant NPCs expressing the negative charge probe RPRE-RFP and CL-GFP show RPRE-RFP translocation from a primarily plasma membrane localization in isogenic-corrected hNs to a punctate intracellular localization in A53T hNs in tight cellular localization with CL-GFP (d). Quantification of percent total cell number with colocalized RPRE-RFP and CL-GFP (e). Data represent mean ± s.e.m. **P < 0.0001 by t-test, n = 3 coverslips over 2 independent differentiations, DIV: 14; scale bar: 10 μm. f, g Fluorescence resonance energy transfer (FRET) from CL-GFP to RPRE-RFP in hiPSC-derived A53T and corrected hNs or hESC-derived WT, A53T and E46K hNs was assessed (f) and mean FRET intensity was quantified (g). Data represent mean ± s.e.m. **P < 0.01 by ANOVA followed by Tukey’s post hoc test, for hiPSCs, n = 6, for hESCs, n = 8 coverslips over two independent differentiations, DIV: 14. Scale bar: 10 µm. h Mitochondria were isolated from the SNpc of 6-month-old WT and A53T transgenic animals and labeled with Annexin V to measure the abundance of cardiolipin on the mitochondrial surface. Controls were rotenone-exposed WT samples. Animals were not randomized, nor was the analysis blinded. Data represent mean ± s.e.m. **P < 0.01 by ANOVA followed by Dunnett post hoc test, n = 4. Clipart was obtained at clker.com Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/s41467-018-03241-9), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Nestin by Immunohistochemistry

Detection of Human Nestin by Immunohistochemistry

MET and TMZ effects in in vivo glioma models(A) Variation of U251 tumor volume measured at contrast-enhanced T1 MRI after treatment expressed as percentage of volume change. Error bars indicate SEM. (B) U251 derived orthotopic brain tumors from vehicle (CTRL), TMZ or TMZ/metformin (Combo) treated mice were analyzed for Ki67, Nestin and CD133 markers by immunohistochemistry. No difference between the tumor edge and bulk areas in IHC staining intensities or percentages of positive cells was observed for any of the investigated markers. H&E was performed for morphological evaluation of the tumors. Representative images are shown with original magnification x200. (C) Variation of T98G tumor volume measured using caliper after treatment expressed as percentage of volume variation. In T98G xenograft model already at 7 days, the combined treatment significantly slowed down tumor growth compared to control (p value = 0.005), moreover a transient reduction was observed with MET (p value = 0.03). After 3 weeks, only Combo-treated mice displayed tumor smaller than vehicle (p value = 0.04). Error bars indicate SEM. (D) T98G tumors from vehicle (CTRL), TMZ, MET or TMZ/metformin (Combo) treated mice, were analyzed for Ki67, Nestin and CD133 markers expression by immunohistochemistry. No difference between the tumor edge and bulk areas in IHC staining intensities or percentages of positive cells was observed for any of the investigated markers. Hematoxylin and eosin staining (H&E) was performed for morphological evaluation of the tumors. Representative images are shown with Original magnification x200. Image collected and cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.23028), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Nestin by Immunohistochemistry

Detection of Human Nestin by Immunohistochemistry

MET and TMZ effects in in vivo glioma models(A) Variation of U251 tumor volume measured at contrast-enhanced T1 MRI after treatment expressed as percentage of volume change. Error bars indicate SEM. (B) U251 derived orthotopic brain tumors from vehicle (CTRL), TMZ or TMZ/metformin (Combo) treated mice were analyzed for Ki67, Nestin and CD133 markers by immunohistochemistry. No difference between the tumor edge and bulk areas in IHC staining intensities or percentages of positive cells was observed for any of the investigated markers. H&E was performed for morphological evaluation of the tumors. Representative images are shown with original magnification x200. (C) Variation of T98G tumor volume measured using caliper after treatment expressed as percentage of volume variation. In T98G xenograft model already at 7 days, the combined treatment significantly slowed down tumor growth compared to control (p value = 0.005), moreover a transient reduction was observed with MET (p value = 0.03). After 3 weeks, only Combo-treated mice displayed tumor smaller than vehicle (p value = 0.04). Error bars indicate SEM. (D) T98G tumors from vehicle (CTRL), TMZ, MET or TMZ/metformin (Combo) treated mice, were analyzed for Ki67, Nestin and CD133 markers expression by immunohistochemistry. No difference between the tumor edge and bulk areas in IHC staining intensities or percentages of positive cells was observed for any of the investigated markers. Hematoxylin and eosin staining (H&E) was performed for morphological evaluation of the tumors. Representative images are shown with Original magnification x200. Image collected and cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.23028), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Rat Nestin by Immunocytochemistry/Immunofluorescence

Detection of Rat Nestin by Immunocytochemistry/Immunofluorescence

Neural stem cells could proliferate and differentiate into neurons and astrocytes.(A) Representative photomicrograph of neurospheres in culture. (B) Immunocytochemical staining of purified NSCs with Nestin. (C) Immunocytochemical staining of purified protoplasmic astrocytes with GFAP. (D) Immunocytochemical staining of purified neurons with beta -tubulin-III. (E) Nucleus staining of differentiated cells from NSCs with DAPI. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26431046), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Rat Nestin by Western Blot

Detection of Rat Nestin by Western Blot

miR–381 promoted neural stem cells proliferation.(A) The expression of miR–381 was measured by qRT-PCR. (B) CCK–8 was performed to detect the neural stem cells proliferation. (C) The mRNA expression of nestin was detected by qRT-PCR. (D) The protein expression of nestin was measured by Western blot. **p<0.01 and ***p<0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26431046), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Nestin by Immunohistochemistry

Detection of Mouse Nestin by Immunohistochemistry

Dissimilar tumorigenesis in the in vivo animal model.(a) Kaplan-Meier survival plots; 3 mice were used for cells of each clone. (b) Representative brain tumors of NOD-SCID mice harboring xenografts of the clones #2 (upper left), #3 (lower left), #4 (upper right) and #5 (lower right); H.E. staining, arrows: tumor, scale bar = 5 mm. (c) A representative xenograft of the clone #4; nestin-positive cells infiltrated the contralateral hemisphere (arrows). Scale bars = 5 mm (left), 1 mm (middle), 50 μm (right). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25623281), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Human Nestin Antibody by Western Blot

Detection of Mouse Human Nestin Antibody by Western Blot

Cardiolipin is externalized to the mitochondrial surface in alpha -syn mutant hNs and transgenic mice in response to alpha -syn accumulation. a Western analysis of lysates from corrected and A53T cells at DIV 14 and DIV 60 labeled for total alpha -syn, PS129, TH (DA neuronal marker), nestin (NPC marker) or GAPDH show that levels of PS129-modified alpha -syn are elevated in A53T hNs at DIV 14 and remain elevated at DIV 60 relative to corrected cells. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29483518), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Nestin by Western Blot

Detection of Human Nestin by Western Blot

Validation of the phosphorylation status regarding the representative phosphoproteins by western blot analyses.Glioblastoma initiating cells were treated with 20 ng/ml EGF for 15 min and subjected to immunoblotting with the corresponding antibodies. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/22912867), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Nestin by Western Blot

Detection of Nestin by Western Blot

VAMP5 KO doesn’t impair the stemness of ESCs.a–d Representative western blot images of VAMP5 KO potentially impair the related markers expression in hESCs (a) and also after differentiated into mesoderm (MES) (b), endoderm (END), (c) and ectoderm (ECT) (d). Data are represented as mean ± SEM, n = 3 biological replicates in each group. Student’s two tailed t test was used. No adjustments for multiple comparisons were made, as only two groups were compared. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. e PCA of the transcriptomic comparison of VAMP5 WT and KO in hESCs and followed differentiated three germ layers, n = 3 in each cell type. f Representative heatmap of hESCs and three germ layers related markers comparison of VAMP5 WT and KO in hESCs and followed differentiated three germ layers. g Immunofluorescence staining for lineage differentiation from VAMP5 WT and KO hESCs to mesoderm derived smooth muscles cells (markers of alpha -SMA and Transgelin), endoderm derived alveolar type II cells (markers of EpCAM and SP-B), and ectoderm derived neuron cells ( beta 3 tubulin and MAP2). The results are representative of three independent experiments. Scale bars, 50 μm. KO, knock-out; WT, wild-type. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40624080), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Nestin Antibody

Application
Recommended Usage

CyTOF-reported

This clone has been commercially reported for use in CyTOF®. Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human neural progenitor cells

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: A172 human glioblastoma cells fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).

Reviewed Applications

Read 7 reviews rated 4.3 using MAB1259 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Nestin

Nestin is a class VI intermediate filament protein (1, 2) that is expressed in stem cells of the central nervous system (CNS) (3) but not in mature CNS cells (4). Nestin expression is used extensively as a marker for CNS stem cells in the developing nervous system and in vitro cultured cells (5-10). Its transient expression is a critical step in the neural differentiation pathway (2). Nestin is also expressed in non-neural stem cell populations, such as pancreatic islet progenitors (11-13) and hematopoietic progenitors (14).

References

  1. Hockfield, S. and R.D. McKay (1985) J. Neurosci. 5:3310.
  2. Lendahl, U. et al. (1990) Cell 60:585.
  3. Frederiksen, K. and R.D. McKay (1988) J. Neurosci. 8:1144.
  4. Tohyama, T. et. al. (1992) Lab. Invest. 66:303.
  5. Uchida, N. et al. (2000) Proc. Natl. Acad.Sci. USA 97:14720.
  6. Frederiksen, K. et al. (1988) Neuron 1:439.
  7. Cattaneo, E. et al. (1990) Nature 347:762.
  8. Reynolds, B.A. and S. Weiss (1992) Science 255:1707.
  9. Rietze, R.L. et al. (2001) Nature 412:736.
  10. Carpenter, M.K. et al. (2001) Exp. Neurol 172:383.
  11. Zulewski, H. et al. (2001) Diabetes 50:521.
  12. Lumelsky, N. et al. (2001) Science 292:1389.
  13. Lechner, A. et al. (2002) Biochem.Biophys. Res. Commun. 293:670.
  14. Shih, C.C. et al. (2001) Blood 98:2412.

Alternate Names

NES

Entrez Gene IDs

10763 (Human); 18008 (Mouse); 25491 (Rat)

Gene Symbol

NES

Additional Nestin Products

Product Documents for Human Nestin Antibody

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Product Specific Notices for Human Nestin Antibody

For research use only

Citations for Human Nestin Antibody

Customer Reviews for Human Nestin Antibody (7)

4.3 out of 5
7 Customer Ratings
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Showing  1 - 5 of 7 reviews Showing All
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  • Human Nestin Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Neuroblastoma cells
    Species: Human
    Verified Customer | Posted 06/17/2022
    Human Nestin Antibody MAB1259
  • Human Nestin Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: Neural stem cells
    Species: Human
    Verified Customer | Posted 02/10/2022
    Human neural stem cells
    Human Nestin Antibody MAB1259
  • Human Nestin Antibody
    Name: Anonymous
    Application: Immunofluorescence - paraffin
    Sample Tested: human glioblastoma
    Species: Human
    Verified Customer | Posted 03/09/2019
    Glioblastoma multiforme stained with anti-nestin antibody 1:100
    pH9 heat induced antigen retrieval
    Human Nestin Antibody MAB1259
  • Human Nestin Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HeLa cells
    Species: Human
    Verified Customer | Posted 10/26/2015
    Specificity: Reasonably specific<br />Sensitivity: Reasonably sensitive<br />Buffer: 1% BSA + 0.1% Triton X-100 in PBS<br />Dilution: 1/100
    Human Nestin Antibody MAB1259
  • Human Nestin Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Human cell line
    Species: Human
    Verified Customer | Posted 10/26/2015
    Specificity: Specific<br />Sensitivity: Reasonably sensitive<br />Buffer: 1% BSA + 0.3% Triton X-100 in PBS<br />Dilution: 1/100
    Human Nestin Antibody MAB1259
  • Human Nestin Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Cell Lysates
    Species: Human
    Verified Customer | Posted 10/26/2015
    Specificity: Reasonably specific<br />Sensitivity: Reasonably sensitive<br />Buffer: 5% non-fat milk in TBST<br />Dilution: 1/1000
    Human Nestin Antibody MAB1259
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 24213561
    Species: Human
    Verified Customer | Posted 02/04/2015

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