Anti-Nestin antibody [SP103] ab105389 is a rabbit monoclonal antibody that is used in Nestin western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with NES knockout cell line validation
- Antibody clone SP103 is cited in over 90 publications
- One antibody for all your Nestin staining, use in Nestin western blotting, IHC, immunofluorescence and flow cytometry
IgG
Rabbit
pH: 7.6
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Liquid
Monoclonal
Flow Cyt (Intra) | WB | IHC-P | ICC/IF | IHC-Fr | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Dog | Predicted | Predicted | Predicted | Predicted | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/50 | Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Dog | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Dog | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes Antigen retrieval: Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min. |
Species | Dilution info | Notes |
---|---|---|
Species Dog | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/200 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Dog | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 5 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Dog | Dilution info - | Notes - |
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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 (By similarity).
Nbla00170, Nbla00170, NES, Nestin
Anti-Nestin antibody [SP103] ab105389 is a rabbit monoclonal antibody that is used in Nestin western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with NES knockout cell line validation
- Antibody clone SP103 is cited in over 90 publications
- One antibody for all your Nestin staining, use in Nestin western blotting, IHC, immunofluorescence and flow cytometry
IgG
Rabbit
pH: 7.6
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Liquid
Monoclonal
SP103
Affinity purification Protein A/G
Purified from TCS by protein A/G.
Blue Ice
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
This supplementary information is collated from multiple sources and compiled automatically.
Nestin also known as NES is an intermediate filament protein with a molecular weight of about 220 kDa. Nestin features prominently in cells related to the nervous system including neural stem cells and it serves as a structural component in these cells. Nestin also appears in substantial amounts during the development of the nervous system and is important for cell structural organization. It is often used as a Nestin marker in research particularly with Nestin antibodies for identifying the presence of neural stem cells and tracking their developmental course. Researchers frequently utilize techniques like Nestin IHC (immunohistochemistry) and Nestin western blot to stain and detect Nestin allowing detailed investigations into its expression in various tissues.
Nestin plays a significant role in the dynamic remodeling of the cytoskeleton. It does not act as a solitary element but forms part of a larger protein complex involved in maintaining and organizing cellular structure. This property grants cells flexibility in shape and resilience during the rapid changes in development and specialization. The role of Nestin in the cellular architecture allows it to contribute to processes such as cell migration and proliferation critical during tissue development and repair.
Nestin integrates into networks that regulate stem cell behavior and neurogenesis. It tightly interacts within the pathways involving cell cycle regulation and differentiation. For example the Wnt and Notch signaling pathways modulate its expression to influence neural stem cell fate and patterning during development. Protein interactions between Nestin and other cytoskeletal elements like vimentin facilitate its roles in these pathways adapting the cellular environment for specific developmental activities.
Nestin associates with various pathological conditions including brain tumors and muscular dystrophies. Its expression is aberrant in many forms of gliomas where it often signifies a higher grade of malignancy. Within these tumors Nestin connects to oncogenic signaling pathways involving proteins such as the epidermal growth factor receptor (EGFR). Moreover in muscular dystrophies Nestin's expression in regenerating muscle fibers illustrates its involvement in cellular repair mechanisms potentially interacting with dystrophin a protein important for muscle integrity. Research on anti-Nestin antibodies continues to clarify its role in these diseases potentially offering diagnostic or therapeutic insights.
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We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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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)
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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Nestin antibody [SP103] (ab105389) at 1/100 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: NES knockout HAP1 whole cell lysate at 20 µg
Predicted band size: 177 kDa
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 Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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.
Formalin-fixed, paraffin-embedded human endometrium tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
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
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.
All lanes: Western blot - Anti-Nestin antibody [SP103] (ab105389) at 1/100 dilution
All lanes: HeLa cell lysate
Predicted band size: 177 kDa
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) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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.
Formalin-fixed, paraffin-embedded human colon tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Formalin-fixed, paraffin-embedded human prostate tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Formalin-fixed, paraffin-embedded human lung tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Formalin-fixed, paraffin-embedded human breast tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Formalin-fixed, paraffin-embedded human skin tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Formalin-fixed, paraffin-embedded human cerebellum tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Formalin-fixed, paraffin-embedded human cervix tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Formalin-fixed, paraffin-embedded human kidney tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
Formalin-fixed, paraffin-embedded human ovary tissue stained for Nestin using ab105389 at 1/100 dilution in immunohistochemical analysis.
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