Nestin Antibody (10C2) - BSA Free

Novus Biologicals | Catalog # NB300-266

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rabbit (Negative), Rat (Negative)

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 10C2

Format

BSA Free
View all Nestin Primary Antibodies »

Product Specifications

Immunogen

A fragment of the human Nestin protein corresponding to residues 1464-1614. [UniProt P48681]

Reactivity Notes

This antibody does not cross-react with rat or rabbit.

Localization

Cytoplasmic

Marker

Neural Stem Cell Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for Nestin Antibody (10C2) - BSA Free

Immunohistochemistry: Nestin Antibody (10C2) - BSA Free [NB300-266]

Immunohistochemistry: Nestin Antibody (10C2) - BSA Free [NB300-266]

Nestin-Antibody-10C2-Immunohistochemistry-NB300-266-img0020.jpg
Western Blot: Nestin Antibody (10C2)BSA Free [NB300-266]

Western Blot: Nestin Antibody (10C2)BSA Free [NB300-266]

Western Blot: Nestin Antibody (10C2) [NB300-266] - WB analysis of 5 ug lysate from (1) Rat's total brain tissue, and (2) human CNS progenitor cells using Nestin antibody (clone 10C2) at 1:1000 dilution. The antibody did not react with rat, whereas, in lysate from human samples, it generated a doublet band representing the phosphorylated and the non-phosphorylated forms of Nestin protein.
Immunocytochemistry/ Immunofluorescence: Nestin Antibody (10C2) - BSA Free [NB300-266]

Immunocytochemistry/ Immunofluorescence: Nestin Antibody (10C2) - BSA Free [NB300-266]

Immunocytochemistry/Immunofluorescence: Nestin Antibody (10C2) [NB300-266] - Analysis of formalin fixed, 0.1% Triton X-100 permeabilized F9 cells (Mouse teratocarcinoma) using Nestin antibody (clone 10C2) at 1:50 dilution. The primary antibody was detected using Alexa Fluor 488 labeled donkey anti-mouse IgG (H+L) cross-adsorbed secondary antibody (green) and nuclei were counterstained with DAPI (blue). An isotype control set was inclubated with mouse IgG1 isotype control antibody and the above mentioned secondary antibody. Nestin antibody (clone 10C2) generated a specific membrane-cytoplasmic staining of Nestin while the isotype control samples did not show any staining under same assay conditions.
Immunohistochemistry-Paraffin: Nestin Antibody (10C2) - BSA Free [NB300-266]

Immunohistochemistry-Paraffin: Nestin Antibody (10C2) - BSA Free [NB300-266]

Immunohistochemistry-Paraffin: Nestin Antibody (10C2) [NB300-266] - Nestin was detected in immersion fixed paraffin-embedded sections of human kidney using anti-human mouse monoclonal antibody (Catalog # NB300-266) at 1:500 dilution overnight at 4C. Tissue was stained using the VisuCyte anti-mouse HRP polymer detection reagent (Catalog # VC001) with DAB chromogen (brown) and counterstained with hematoxylin (blue).
Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand.
Flow Cytometry: Nestin Antibody (10C2) - BSA Free [NB300-266]

Flow Cytometry: Nestin Antibody (10C2) - BSA Free [NB300-266]

Flow Cytometry: Nestin Antibody (10C2) [NB300-266] - Intracellular flow cytometric staining of 1 x 10^6 CHO (A) and HEK-293 (B) cells using Nestin antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1 x 10^6 cells was used.
Immunocytochemistry/ Immunofluorescence: Nestin Antibody (10C2) - BSA Free [NB300-266]

Immunocytochemistry/ Immunofluorescence: Nestin Antibody (10C2) - BSA Free [NB300-266]

Immunocytochemistry/Immunofluorescence: Nestin Antibody (10C2) [NB300-266] - Visualization of Nestin in PC-3 cells using NB300-266. Courtesy of Dolnick Lab, Roswell Park Cancer Institute.
Flow Cytometry: Nestin Antibody (10C2) - BSA Free [NB300-266]

Flow Cytometry: Nestin Antibody (10C2) - BSA Free [NB300-266]

Flow Cytometry: Nestin Antibody (10C2) [NB300-266] - An intracellular stain was performed on SH-SY5Y cells with Nestin [10C2] Antibody NB300-266C (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 650.
Flow Cytometry: Nestin Antibody (10C2) - BSA Free [NB300-266]

Flow Cytometry: Nestin Antibody (10C2) - BSA Free [NB300-266]

Flow Cytometry: Nestin Antibody (10C2) [NB300-266] - An intracellular stain was performed on HeLa cells with Nestin Antibody (10C2)NB300-266PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.
Nestin Antibody (10C2) - BSA Free

Immunohistochemistry: Nestin Antibody (10C2) - BSA Free [NB300-266] -

nb300-266_mouse-monoclonal-nestin-antibody-10c2-20920231412574.jpg
Nestin Antibody (10C2) - BSA Free

Immunohistochemistry: Nestin Antibody (10C2) - BSA Free [NB300-266] -

Immunohistochemistry: Nestin Antibody (10C2) - BSA Free [NB300-266] - Regorafenib inhibits tumour growth & vascularisation in xenograft mice. GC1 were implanted into the right forebrain of mice. The mice then received daily oral vehicle or regorafenib at 30 mg/kg for 45 days. (A) Representative photographs of mice brain tumours treated with regorafenib (right panel) or without regorafenib (left panel) stained by hemalun–eosin (median panel) or with nestin antibody (upper & lower panel). The upper panel shows whole brains of mice stained with nestin antibody. The tumour areas were circled in blue, & each surface was measured in mm2. The red rectangles indicate the zone of the tumour shown in median (hemalun–eosin staining) & lower (nestin IHC) panel. Scale bars, upper panel 2.5 mm, median & lower panel 100 µm. The graph shows the tumour size in mm2 (expressed as the mean ± SEM of 4 mice). (B) Representative immunohistochemistry photographs of CD31+ vessels in brain tumour areas of 3 different mice treated with regorafenib (right panel) or without regorafenib (control, left panel). Arrows indicate CD31+ blood vessels. Scale bars, 20 µm. The graph shows the number of CD31+ vessels in tumours per mm2 (expressed as mean ± SEM of 4 mice). * p < 0.05. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35326702), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Nestin Antibody (10C2) - BSA Free

Nestin (10C2) in SJCRH30 Human Cell Line.

Nestin (10C2) was detected in immersion fixed SJCRH30 human Rhabdomyosarcoma cell line with Mouse anti-Nestin (10C2) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB300-266AF488) (green) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (dark blue). Cells were imaged using a 100X objective and digitally deconvolved.
Nestin Antibody (10C2) - BSA Free

Detection of Nestin (10C2) in SJCRH30 Human Cell Line by Flow Cytometry.

An intracellular stain was performed on SJCRH30 human Rhabdomyosarcoma cell line with Mouse anti-Nestin (10C2) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB300-266AF488, blue histogram) or matched control antibody (orange histogram) at 5 µg/mL for 30 minutes at RT.
Nestin Antibody (10C2) - BSA Free

Nestin (10C2) in SJCRH30 Human Cell Line.

Nestin (10C2) was detected in immersion fixed SJCRH30 human Rhabdomyosarcoma cell line with Mouse anti-Nestin (10C2) Protein-G purified Monoclonal Antibody conjugated to DyLight 650 (Catalog # NB300-266C) (light blue) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (dark blue). Cells were imaged using a 100X objective and digitally deconvolved.
Nestin Antibody (10C2) - BSA Free

Nestin (10C2) in SJCRH30 Human Cell Line.

Nestin (10C2) was detected in immersion fixed SJCRH30 human Rhabdomyosarcoma cell line using Mouse anti-Nestin (10C2) Protein G-purified Monoclonal Antibody (Catalog # NB300-266) at 2.0 µg/mL overnight at 4C. Cells were stained using DyLight 488-conjugated Anti-Mouse IgG (H+L) Cross-Absorbed Secondary Antibody (green), and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

Applications for Nestin Antibody (10C2) - BSA Free

Application
Recommended Usage

Flow Cytometry

1 ug per million cells

Immunocytochemistry/ Immunofluorescence

1:50 - 1:200

Immunohistochemistry

1:100 - 1:200

Immunohistochemistry-Frozen

1:100 - 1:200

Immunohistochemistry-Paraffin

1:100 - 1:200

Western Blot

1:1000 - 1:5000
Application Notes
This Nestin (10C2) antibody is useful for Immunohistochemistry on formaldehyde fixed cultured cells and formalin fixed paraffin-embedded tissue sections. In Western blot a doublet band is seen at ~220-240 kDa. The investigator should determine the optimal working dilution for a specific application. This antibody is CyTOF ready.

Reviewed Applications

Read 2 reviews rated 5 using NB300-266 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Nestin

Nestin is a Class VI intermediate filament expressed in the developing central nervous system (CNS) in early embryonic neuroepithelial stem cells. This protein has been widely used as a predominant marker for stem / progenitor cells, glioma cells, and tumor endothelial cells. Furthermore, it is a superior angiogenic marker to evaluate neovascularity of endothelial cells in tumor.

Alternate Names

NES

Gene Symbol

NES

Additional Nestin Products

Product Documents for Nestin Antibody (10C2) - BSA Free

Certificate of Analysis

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Product Specific Notices for Nestin Antibody (10C2) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Nestin Antibody (10C2) - BSA Free

Customer Reviews for Nestin Antibody (10C2) - BSA Free (2)

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  • Nestin Antibody (10C2)
    Name: John H
    Application: Western Blot
    Sample Tested: Primary glioblastoma cells - whole cell lysate and Primary glioblastoma cells
    Species: Human
    Verified Customer | Posted 07/13/2017
    40 ug whole cell lysate Blocking - 5% Milk TBST Antibody - 1:2000 overnight at 4C
  • Name: Shama Mirza
    Application: Western Blot
    Sample Tested:
    Species: Human
    Verified Customer | Posted 08/14/2014

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FAQs for Nestin Antibody (10C2) - BSA Free

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  • Q:  Why do you show the reactivity of mouse as “negative”? Could you provide me any further information for this description?

    A: Upon review of the publication, it seems that the authors were testing nestin-GFP-transgenic mice. This may have affected the results versus endogenous detection, which we have performed in our lab. Per our testing, we have found that this clone usually does not detect the mouse protein in endogenous samples. That is why we list this species as negative on our datasheet. Additionally, the immunogen for this antibody only shares 45% homology to the mouse sequence.

  • Q: We are interested in your item, NB300-266 and is it possible to provide me the % of predicted cross-reactivity with pig based on sequence homology?

    A: The immunogen of NB300-266 is a fragment of the human Nestin protein corresponding to residues 1464-1614. [UniProt P48681] and the BLAST results of this sequence may be seen at : Uniprot Site This sequence shows ~64% or less similarity with some un-characterized proteins in pig some of which might be NES. The alignment results of your mentioned sequence with the immunogen sequence of this product may be seen at: Uniprot Site There are several amino acids which are similar to immunogen of NB300-266, however, considering it being a monoclonal, I am not very hopeful that it will cross-react with the putative NES in pig. Accordingly, we will not guarantee it for Pig samples.

  • Q:  Why do you show the reactivity of mouse as “negative”? Could you provide me any further information for this description?

    A: Upon review of the publication, it seems that the authors were testing nestin-GFP-transgenic mice. This may have affected the results versus endogenous detection, which we have performed in our lab. Per our testing, we have found that this clone usually does not detect the mouse protein in endogenous samples. That is why we list this species as negative on our datasheet. Additionally, the immunogen for this antibody only shares 45% homology to the mouse sequence.

  • Q: We are interested in your item, NB300-266 and is it possible to provide me the % of predicted cross-reactivity with pig based on sequence homology?

    A: The immunogen of NB300-266 is a fragment of the human Nestin protein corresponding to residues 1464-1614. [UniProt P48681] and the BLAST results of this sequence may be seen at : Uniprot Site This sequence shows ~64% or less similarity with some un-characterized proteins in pig some of which might be NES. The alignment results of your mentioned sequence with the immunogen sequence of this product may be seen at: Uniprot Site There are several amino acids which are similar to immunogen of NB300-266, however, considering it being a monoclonal, I am not very hopeful that it will cross-react with the putative NES in pig. Accordingly, we will not guarantee it for Pig samples.

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