Nestin Antibody - BSA Free

Novus Biologicals | Catalog # NB100-1604

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Baboon, Monkey

Cited:

Human, Mouse, Rat, Baboon

Applications

Validated:

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

Cited:

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

Label

Unconjugated

Antibody Source

Polyclonal Chicken IgY

Format

BSA Free
View all Nestin Primary Antibodies »

Product Specifications

Immunogen

Chickens were immunized with three synthetic peptide/keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of mouse Nestin (NM_NP_057910), but are shared with the rat (NM_012987, NCBI) protein sequence.

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 19481056). Baboon reactivity reported in scientific literature (PMID: 30657788). Monkey reactivity reported in scientific literature (PMID: 30657788). Human reactivity reported in scientific literature (PMID: 33023162).

Localization

Cytoplasmic

Marker

Neural Stem Cell Marker

Clonality

Polyclonal

Host

Chicken

Isotype

IgY

Theoretical MW

250 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Nestin Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604]

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604]

Nestin-Antibody-Immunocytochemistry-Immunofluorescence-NB100-1604-img0008.jpg
Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604]

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604]

Nestin-Antibody-Immunocytochemistry-Immunofluorescence-NB100-1604-img0007.jpg
Immunohistochemistry-Frozen: Nestin Antibody [NB100-1604]

Immunohistochemistry-Frozen: Nestin Antibody [NB100-1604]

Immunohistochemistry-Frozen: Nestin Antibody [NB100-1604] - Analysis of Nestin in frozen mouse tissue using anti-Nestin antibody. IHC-Fr image submitted by a verified customer review.
Nestin Antibody Western Blot: Nestin Antibody [NB100-1604]

Western Blot: Nestin Antibody [NB100-1604]

20 ug of total protein from adult mouse brain using NB100-1604 (1:2000)
Nestin Antibody Immunohistochemistry: Nestin Antibody [NB100-1604]

Immunohistochemistry: Nestin Antibody [NB100-1604]

Photomicrograph of a tissue section through the periventricular zone of an e16 mouse brain (4.0% paraformaldehyde-fixed). Green is Nestin immunoreactivity (1:1000 dilution). Blue is DAPI nuclear staining.
Nestin Antibody

Immunohistochemistry: Nestin Antibody [NB100-1604] -

Representative images of nestin (A) and Tmsb4x (B) staining in the mouse adult wild-type glomerulus visualized by fluorescent microscopy. (C) Merged images showing Tmsb4x (red) and nestin (green) staining; areas of colocalization are indicated by arrows. Representative images of Tmsb4x (D) and Cd31 (E) staining in the mouse adult wild-type glomerulus visualized by fluorescent microscopy. (F) Merged images showing Tmsb4x (green) and Cd31 (red) staining. Bar = 20 μm.
Nestin Antibody

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604] -

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604] - In vitro proliferation & differentiation of ahMNCs-Clump II. (A) ahMNCs-Clump II were propagated in the culture condition for ahMNCs. The accumulated number of cells of ahMNCs-Clump II (NS18-007TL & 008TL) in comparison with ahMNCs established using previous culture methods (NS14-010TL, NS14-011TL, & NS15-001TL). Days to 1 × 109 cells are indicated. (B) Morphologies of ahMNCs-Clump II under expansion processes are illustrated until third in vitro passage (P3). Scale bar = 50 μm. (C) After in vitro differentiation, immunofluorescence was applied to ahMNCs-Clump II. Nestin for NSCs, MAP2 & Tuj1 for neurons, & GFAP for astrocytes. Undiffer = before in vitro differentiation; Differ = after in vitro differentiation. Scale bar = 10 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30380605), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Nestin Antibody

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604] -

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604] - Representative images of nestin (A) & Tmsb4x (B) staining in the mouse adult wild-type glomerulus visualized by fluorescent microscopy. (C) Merged images showing Tmsb4x (red) & nestin (green) staining; areas of colocalization are indicated by arrows. Representative images of Tmsb4x (D) & Cd31 (E) staining in the mouse adult wild-type glomerulus visualized by fluorescent microscopy. (F) Merged images showing Tmsb4x (green) & Cd31 (red) staining. Bar = 20 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27575556), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Nestin Antibody

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604] -

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604] - Dopaminergic neuron derivation from H9 stem cells & hIPSCs.Scheme showing the different steps of the DA neuron derivation protocol (A). Immunolabelings of cultured H9 (top panels) & WT hiPSCs (bottom panels) for different NSCs markers, Sox1, Pax6 & Nestin (B). Immunolabeling of cultured H9 (top panels) & WT hiPSCs (bottom panels) after 3 days of DA progenitor induction for progenitor markers Lmx1A & FoxA2 (C) & after 7 days of induction for immature neurons beta III-Tubulin+ & remaining progenitors Sox1+ (D). Immunolabelings of cultured H9 (top panels) & WT hiPSCs (bottom panels) after 21 days of induction for the neuronal marqueur MAP2 & DA neurons Tyrosine Hydroxylase (TH) (E). Analysis of gene expression levels by qRT-PCR for cultured H9 (top panels) & WT hiPSCs at different time points (NSC, 14 days & 21 days) of the culture (FB = fetal brain) (F,G). Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep33377), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Nestin Antibody

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604] -

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604] - Neural Stem Cell derivation from WT & LRRK2 G2019S hiPSCs.Haematoxylin & eosin coloration of teratoma-encapsulated tumors generated by flank injection of WT & G2019S hiPSCs cells in Nod/Scid mice. Representative images of the different germ layers: ectoderm (immature squamous epithelium, neural rosettes), mesoderm (primitive cartilage, muscles, fat) & endoderm (primitive gut like epithelium) (A). Scheme showing the different steps of the DA neuron derivation protocol (B). Immunolabelings of cultured WT & LRRK2 G2019S hIPSCs for different NSCs markers after NSCs induction protocol: Sox1, Nestin, Pax6 & Ki67 (C). Analysis of gene expression levels by qRT-PCR for WT & LRRK2 G2019S cultures at the end of the NSCs induction protocol (D). Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep33377), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Nestin Antibody

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604] -

Immunocytochemistry/ Immunofluorescence: Nestin Antibody [NB100-1604] - PAR1 is expressed by neural stem cells in the sub-ventricular zone (SVZ) of the adult mouse brain. Photomicrographs show immunofluorescence double-labeling for PAR1 with Sox2-positive (A), or PAR1 with Nestin-positive (C) multipotent neural stem cells (NSCs) within the lateral wall of the lateral ventricle (LV). RNAscope was used to identify cells expressing both PAR1 & Sox2 (B), or Nestin (D) RNA in NSCs of the adult SVZ. Arrow indicates an example of a double-labeled cell in each case, with arrowhead indicating a singly labeled cell (Scale bar = 10 μm). Boxed area in B & D is also shown at higher magnification to visualize double-labeled cells. (E) Histogram shows expression of PAR1 RNA was high in NSCs grown as neurospheres (NS), or when plated on poly-L-lysine coated coverslips as monolayers in stem cell media containing EGF & bFGF. PAR1 RNA expression by NSC monolayers decreased by 87% when EGF & bFGF were removed from the media for 7 DIV promoting stem cell differentiation. (F) Withdrawal of EGF & bFGF to induce NSC monolayer differentiation resulted in a parallel decrease in Nestin RNA expression. (**P < 0.01, ***P < 0.001 Students t-test). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29921916), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Nestin Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:1000 - 1:2000

Immunohistochemistry

1:1000 - 1:2000

Immunohistochemistry-Frozen

1:1000 - 1:2000

Immunohistochemistry-Paraffin

1:1000 - 1:2000

Western Blot

1:10000 - 1:20000

Reviewed Applications

Read 5 reviews rated 5 using NB100-1604 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
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  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

10mM PBS (0.9% isotonic, w/v, pH 7.2)

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

0.3 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 in the dark.

Background: Nestin

Nestin is a Class VI intermediate filament expressed in the developing central nervous system 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. Because if it's roles in angiogenisis and tumorgenesis, Nestin antibodies are useful cancer markers.

Although it is often used as a marker for proliferating and migrating cells very little is known about Nestin's function or regulation.

Alternate Names

NES

Entrez Gene IDs

18008 (Mouse)

Gene Symbol

NES

UniProt

Q6P5H2 (Mouse), NP_057910 (Mouse)

Additional Nestin Products

Product Documents for Nestin Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for Nestin Antibody - BSA Free

Chicken products cannot be exported to Canada.

Purification Notes
After repeated injections, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity-purified using a peptide column, and the concentrations of the eluates adjusted to 300 ug/ml. Finally, equal volumes of each of the three affinity-purified anti-peptide antibodies were mixed, and the preparation was filter sterilized.

Storage Notes
Store at 4C in the dark. Under these conditions, the antibodies should have a shelf life of at least 12 months (provided they remain sterile). Do not freeze these antibodies unless you want to store them for longer periods of time. Note, however, that each time an antibody preparation is frozen, about half of its binding activity is lost.

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 - BSA Free

Customer Reviews for Nestin Antibody - BSA Free (5)

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Customer Images

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  • Nestin Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: Neural Cells
    Species: Mouse
    Verified Customer | Posted 07/26/2021
    Expression of Nestin by neural cells in mouse
    Nestin Antibody - BSA Free NB100-1604
  • Nestin Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: fibroblasts
    Species: Mouse
    Verified Customer | Posted 12/16/2020
    Nestin (Green) in fibroblasts
    Nestin Antibody - BSA Free NB100-1604
  • Nestin Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: bEnd.3 mouse endothelioma cell line
    Species: Mouse
    Verified Customer | Posted 05/01/2019
  • Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested:
    Species: Mouse
    Verified Customer | Posted 01/09/2015
    Nestin Antibody - BSA Free NB100-1604
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 23741377
    Species: Mouse
    Verified Customer | Posted 12/12/2014

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Protocols

View specific protocols for Nestin Antibody - BSA Free (NB100-1604):


Citrate Buffer Antigen Retrieval Protocol



Background: Formaldehyde fixation (2% or 4%, or as a component of 10% formalin) produces protein cross-links in tissues that tends to interfere with antibody penetration. This seems to be particularly true of paraffin- embedded formaldehyde-fixed tissue. Since chicken IgY antibodies are larger than rabbit or mouse IgG's, "extra steps" may be necessary to compensate for their larger size.

The citrate-based "antigen retrieval" protocol outlined below has been shown to improve chicken IgY antibody penetration into 4% formalde- hyde-fixed paraffin-embedded sections, and can increase the degree and intensity of immunoreactivity and immunostaining.


Reagents (NOTE: You can use either the Sodium Citrate or Citric Acid Buffers in step #3, below)

"Sodium Citrate Buffer" (10mM Sodium Citrate, 0.05% Tween 20, pH 6.0)

Weigh out 2.94 grams of trisodium citrate (dihydrate). Dissolve in approximately
900 mls of deionized, distilled water. Adjust the pH to 6.00 with 1.0 N HCl. Add
0.5 ml of Tween-20. Mix. Bring up the volume to 1.0 litres with water. Store this solution at room temperature for 3 months or at 4C for longer periods.

"Citric Acid Buffer" (10mM Citric Acid, 0.05% Tween 20, pH 6.0)

Weigh out 1.92 grams of citric acid (anhydrous). Dissolve in approximately 900 mls of deionized, distilled water. Adjust the pH to 6.0 with 1.0 N NaOH. Add
0.5 ml of Tween-20. Mix. Bring up the volume to 1.0 litres with water. Store this solution at room temperature for 3 months or at 4C for longer periods.

"Phosphate-Buffered Saline" [PBS, 10 mM Sodium phosphate-buffered (pH
7.2) isotonic (0.9%, w/v) saline solution] PBS Tween (0.05% Tween 20 in PBS)
Ethanol (80%, 90%, 95%, 100%) diluted with water

Xylene



Procedure (for use with paraffin-embedded sections):

1 Deparaffinize tissue sections in 2 changes of xylene (5 minutes each).

2. Hydrate in 2 changes of 100% ethanol (3 minutes each), 95% ethanol (1 minute),
90% ethanol (1 minute), 80% ethanol (1 minute). Rinse in distilled water.

3. Pre-heat steamer or water bath with staining dish containing either Sodium
Citrate Buffer or Citrate Buffer. Wait until temperature reaches 95-100 degrees C.

NOTE: Microwave or pressure cooker can be used as an alternative as a heating source.

4. Immerse slides in the staining dish. Place the lid loosely on the staining dish and incubate for 20-40 minutes (optimal incubation times will vary).

5. Remove the staining dish, and allow it to cool to room temperature (for 20 minutes or so).

6. Rinse sections in PBS Tween twice for 2 minutes each time.

NOTE: The remainder of this protocol is meant to be a suggestion, and can be substituted with your regular immunostaining protocol.

7. Block sections for 30 minutes with Blocking buffer diluted 1:10 with water.

8. Incubate sections with primary antibody at appropriate dilution in antibody dilution buffer overnight at 4 degrees C. Since chicken IgY antibodies are larger than mammalian IgG's, this overnight incubation allows more time for antibody penetration into tissue sections.

9. Rinse sections with PBS Tween 20 twice for 5 minutes each time.

10. Incubate sections with labeled secondary antibody (see NOTE, below) at appropriate dilution (for one hour at room temperature) in a 1:100 dilution of blocking buffer (diluted in PBS).

11. Rinse with PBS Tween 20 for three times for 5 minutes each time.

NOTE: This protocol may use HRP- or fluorescently-labeled secondary antibodies produced in goats or rabbits.

References:

1. Shi SR, Chaiwun B, Young L, Cote RJ, Taylor CR. (1993). Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections. J Histochem Cytochem 41 (11): 1599-1604.

2. Kanai K, Nunoya T, Shibuya K, Nakamura T, Tajima M (1998). Variations in effectiveness of antigen retrieval pretreatments for diagnostic immunohistochemistry. Res Vet Sci 64 (1): 57-
61.

3. Brown RW, Chirala R. (1995). Utility of microwave-citrate antigen retrieval in diagnostic immunohistochemistry. Mod Pathol 8 (5): 515-20.

4. Morgan JM, Navabi H, Schmid KW, Jasani B (1994). Possible role of tissue-bound calcium ions in citrate-mediated high-temperature antigen retrieval. J Pathol 174 (4): 301-7.


5. Pellicer EM, Sundblad A (1994). Antigen retrieval by microwave oven with buffer of citric acid.
Medicina (B Aires). 54 (2): 129-32.

6. Shi SR, Chaiwun B, Young L, Cote RJ, Taylor CR (1993). Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections. J Histochem Cytochem 41 (11): 1599-604.

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FAQs for Nestin Antibody - BSA Free

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  • Q: Is this antibody from serum or egg? Is it IgG or IgY antibody? Which secondary antibody we should use?

    A: This antibody was purified from egg yolks. This antibody is isotype IgY. You should definitely use IgY secondary with this antibody.

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