Name: Anti-Nestin antibody [SP103]
Brand: RabMAb
SKU: ab105389
Rating: 5 (7 reviews)

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Western blot - Anti-Nestin antibody [SP103] (ab105389)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [SP103] to Nestin
Suitable for: ICC/IF, Flow Cyt, IHC-Fr, WB, IHC-P
Knockout validated
Reacts with: Human

Carrier Free

Product name

Anti-Nestin antibody [SP103]
See all Nestin primary antibodies
Description

Rabbit monoclonal [SP103] to Nestin
Host species

Rabbit
Tested applications

Suitable for: ICC/IF, Flow Cyt, IHC-Fr, WB, IHC-Pmore details
Species reactivity

Reacts with: Human
Predicted to work with: Dog
Immunogen

Synthetic peptide within Human Nestin aa 1600 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P48681
Positive control
- IHC-P: Human kidney, ovary, cervix, breast, colon, endometrium, cerebellum, lung, prostate, skin and human melanoma tissue. WB: HeLa cell lysate. Flow Cyt: SH-SY5Y cells. ICC/IF: SKNSH cells. IHC-Fr: Frozen human kidney tissue sections
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
Storage buffer

pH: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Concentration information loading...
Purity

Protein A/G purified
Purification notes

Purified from TCS by protein A/G.
Clonality

Monoclonal
Clone number

SP103
Isotype

IgG
Research areas

Alternative Versions
- Anti-Nestin antibody [SP103] - BSA and Azide free (ab271870)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- HeLa whole cell lysate (ab29545)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab105389 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
ICC/IF	(2)	1/200.
Flow Cyt		1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-Fr		Use a concentration of 5 µg/ml.
WB	(2)	1/100. Predicted molecular weight: 177 kDa.
IHC-P	(3)	1/100. Antigen retrieval: Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min.

Notes
ICC/IF 1/200.
Flow Cyt 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-Fr Use a concentration of 5 µg/ml.
WB 1/100. Predicted molecular weight: 177 kDa.
IHC-P 1/100. Antigen retrieval: Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min.

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.
Target information above from: UniProt accession P48681 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10763 Human
- Omim: 600915 Human
- SwissProt: P48681 Human
- Unigene: 527971 Human
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

Western blot - Anti-Nestin antibody [SP103] (ab105389)

All lanes : Anti-Nestin antibody [SP103] (ab105389) at 1/100 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : NES knockout HAP1 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 177 kDa

Lanes 1 - 2: Merged signal (red and green). Green - ab105389 observed at 300 kDa. Red - loading control, ab18058, observed at 117 kDa.

ab105389 was shown to specifically react with NES in wild-type HAP1 cells as signal was lost in NES knockout cells. Wild-type and NES knockout samples were subjected to SDS-PAGE. Ab105389 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/100 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-Nestin antibody [SP103] (ab105389)

ICC/IF image of ab105389 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab105389, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)Malchenko, S. et al PLoS One. 2017 Mar 1;12(3):e0173106. doi: 10.1371/journal.pone.0173106. eCollection 2017 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

BTIC markers in our CNS-PNET model tumors

Formalin-fixed, paraffin-embedded mouse primitive neural ectodermal tumor tissue was stained for Nestin using ab105389 at 1/30 dilution in immunohistochemical analysis.

Insert x20 magnification.

TU-tumor; P-parenchyma; V-ventricle; CP-choroid plexus

(From Figure 3C of Malchenko et al)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

Formalin-fixed, paraffin-embedded human endometrium tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Immunohistochemistry (Frozen sections) - Anti-Nestin antibody [SP103] (ab105389)

IHC image of Nestin staining in a section of frozen normal human kidney* performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab105389, 5ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-Nestin antibody [SP103] (ab105389)

Anti-Nestin antibody [SP103] (ab105389) at 1/100 dilution + HeLa cell lysate

Predicted band size: 177 kDa
Flow Cytometry - Anti-Nestin antibody [SP103] (ab105389)

Overlay histogram showing SH-SY5Y cells stained with ab105389 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab105389, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

Formalin-fixed, paraffin-embedded human colon tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

Formalin-fixed, paraffin-embedded human prostate tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

Formalin-fixed, paraffin-embedded human lung tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

Formalin-fixed, paraffin-embedded human breast tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

Formalin-fixed, paraffin-embedded human skin tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

Formalin-fixed, paraffin-embedded human cerebellum tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

Formalin-fixed, paraffin-embedded human cervix tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

Formalin-fixed, paraffin-embedded human kidney tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)

Formalin-fixed, paraffin-embedded human ovary tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [SP103] (ab105389)This image is courtesy of Hongwei Shao by Abreview.

ab105389 staining Nestin in human melanoma tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using Tris EDTA pH 9.0. Samples were then permeabilized using 0.25% Triton X-100, blocked, then incubated with ab105389 at a 1/100 dilution for 18 hours at 4°C. The secondary used was an Alexa-Fluor 594 (red) conjugated donkey anti-rabbit polyclonal used at a 1/100 dilution. (Green) tumor cells, (blue) DAPI.
See Abreview
Anti-Nestin antibody [SP103] (ab105389)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab105389? Please let us know so that we can cite the reference in this datasheet.

ab105389 has been referenced in 46 publications.

Jin C et al. CircRNA EPHB4 modulates stem properties and proliferation of gliomas via sponging miR-637 and up-regulating SOX10. Mol Oncol 15:596-622 (2021). PubMed: 33085838
Xiao Z et al. Pimozide augments bromocriptine lethality in prolactinoma cells and in a xenograft model via the STAT5/cyclin D1 and STAT5/Bcl-xL signaling pathways. Int J Mol Med 47:113-124 (2021). PubMed: 33155660
Solomon E et al. Global transcriptome profile of the developmental principles of in vitro iPSC-to-motor neuron differentiation. BMC Mol Cell Biol 22:13 (2021). PubMed: 33602141
Xiao Z et al. GASC1 promotes glioma progression by enhancing NOTCH1 signaling. Mol Med Rep 23:N/A (2021). PubMed: 33649841
Chen Q et al. Small molecules efficiently reprogram apical papilla stem cells into neuron-like cells. Exp Ther Med 21:546 (2021). PubMed: 33850518

View all Publications for this product

Customer reviews and Q&As

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1-9 of 9 Abreviews or Q&A

Application

Western blot

Loading amount

5 µg

Gel Running Conditions

Reduced Denaturing (10%)

Sample

Human Cell lysate - other (undifferentiated human iPS cells)

Specification

undifferentiated human iPS cells

Blocking step

5% milk in TBST as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Dec 18 2013

Application

Western blot

Sample

Human Cell lysate - whole cell (PANC-1)

Gel Running Conditions

Reduced Denaturing (10%)

Loading amount

50 µg

Specification

PANC-1

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Dr. Carlos Melo

Verified customer

Submitted May 03 2021

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (hiPSC-NSCs (neural stem cells))

Permeabilization

Yes - 0.05% Triton-X 100

Specification

hiPSC-NSCs (neural stem cells)

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Fixative

Formaldehyde

MRS. Alessandra Granata

Verified customer

Submitted Dec 29 2019

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Blocking step

Dako protein block as blocking agent for 10 minute(s) · Concentration: .25% · Temperature: 22°C

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate

Sample

Dog Tissue sections (Fetal tissue)

Specification

Fetal tissue

Permeabilization

Fixative

Formaldehyde

Ms. Erica Sloma

Verified customer

Submitted Aug 21 2014

Application

Immunocytochemistry/ Immunofluorescence

Blocking step

1% serum, 0.1% triton, 0.1% BSA in PBS as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C

Sample

Human Cell (neural progenitor cell derived from human iPSCs)

Specification

neural progenitor cell derived from human iPSCs

Permeabilization

Yes - (see blocking buffer)

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 22 2013

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (kidney)

Specification

kidney

Fixative

Formaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: EDTA pH9, 20 min, 100C

Permeabilization

Abcam user community

Verified customer

Submitted Nov 22 2011

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Melanoma)

Specification

Melanoma

Fixative

Formaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris EDTA pH9.0

Permeabilization

Yes - 0.25% Triton X-100

Blocking step

DakoCytomation X0909 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C

Hongwei Shao

Verified customer

Submitted Nov 09 2011

Question

Inquiry: I ordered the RabMAb Anti-nestin antibody and I tested it using conditions recommended in the spec sheet for ICC/IF (1:200). I used this antibody at the same time as some other nestin antibodies from other companies, so I know the cells (human glioblastoma stem cells) express a lot of nestin. And I was also testing the RabMab anti-GFAP antibody (ab68428) at the same time, and this antibody worked really well. I have since repeated the experiment using different antibody concentrations and still have negative results (I get no signal). I am using the Alexafluor goat anti- rabbit 488 secondary antibody and again I was testing other rabbit antibodies at the same time and it worked at the concentration I used (1:250). My protocol is: 1. Fix in 8% PFA added directly to cells in culture media in a 1:1 ratio so it's diluted to 4% 2. wash 3. Permeabilize with 0.1% Triton-x-100 4. wash 5. Block in 10% goat serum 6. Add primary antibody in blocking solution for 1 hr at RT (I've gone as low as 1:100) 7. wash 8. Add secondary antibody in blocking solution for 1 hr at RT 9. wash 10. Add Hoescht 33342 I tried to be as detailed as possible, but please let me know if you need anymore info about my experiment. Thanks for your help.

Abcam community

Verified customer

Asked on Jan 16 2013

Answer

Thank you for contacting us. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab105389. I would also appreciate if you can confirm some further details:

1) How long did you permeabilize with Triton?

2) Overnight incubation was used in the image on our datasheet. This is usually the best way to get a strong signal in your cells.

Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

Abcam Scientific Support

Answered on Jan 16 2013

Question

I have a question regarding one of you anti-Nestin antibodies ( ab105389 ). On the website it lists this antibody as reacting with human, but doesn't provide any informatin regarding mouse. I looked at a number of other Nestin antibodies and some react with both mouse and human, and some are specific to one or the other (for example a mouse ab that lists as "doesn't react with human"). I was wondering if based on the antigen used for this antibody if it if would or wouldn't react with mouse.

Abcam community

Verified customer

Asked on Dec 12 2012

Answer

Thank you for contacting us.

Unfortunately, alignments of the immunogen with mouse show very poor homology. As such, we cannot guarantee or recommend this product for use in this species. We do offer a number of other anti-Nestin antibodies of which nine are tested and guaranteed in mouse samples. In general, I would recommend Anti-Nestin antibody [2Q178] - Neural Stem Cell Marker (ab6142) as this product is extensively tested, published with and reviewed. However, I would be happy to help find the right product for you based on your needs.

More information about ab6142 may be found at this location: https://www.abcam.com/nestin-antibody-2q178-neural-stem-cell-marker-ab6142.html

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Abcam Scientific Support

Answered on Dec 12 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Purification notes

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Developmental stage

Post-translationalmodifications

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (46)

Customer reviews and Q&As

Western blot abreview for Anti-Nestin antibody [SP103]

Western blot abreview for Anti-Nestin antibody [SP103]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Nestin antibody [SP103]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nestin antibody [SP103]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Nestin antibody [SP103]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nestin antibody [SP103]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nestin antibody [SP103]

Post-translational
modifications