Recombinant Anti-Nestin antibody [EPR1301(2)] (ab176571)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1301(2)] to Nestin
- Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-Nestin antibody [EPR1301(2)]
See all Nestin primary antibodies -
Description
Rabbit monoclonal [EPR1301(2)] to Nestin -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IHC-Pmore details
Unsuitable for: IP or WB -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Nestin aa 550-650 (Cysteine residue). The exact sequence is proprietary.
Database link: P48681 -
Positive control
- IHC-P: Human kidney, spleen and melanoma tissues. ICC/IF: This antibody gave a positive result when used in the following methanol fixed cell lines: SKNSH, U-87 MG Flow Cyt (intra): SH-SY5Y
-
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1301(2) -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab176571 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 |
---|---|---|
Flow Cyt (Intra) |
1/160.
|
|
ICC/IF |
1/50 - 1/100.
For unpurified use at 1/50 - 1/100. |
|
IHC-P |
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
The use of an HRP/AP polymerized secondary antibody is recommended. See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
Notes |
---|
Flow Cyt (Intra)
1/160. |
ICC/IF
1/50 - 1/100. For unpurified use at 1/50 - 1/100. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. The use of an HRP/AP polymerized secondary antibody is recommended. See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
Target
-
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
modificationsConstitutively phosphorylated. This increases during mitosis when the cytoplasmic intermediate filament network is reorganized. - 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
see all
Images
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human spleen tissue sections labeling Nestin with purified ab176571 at 1:1000 dilution (1.65 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling Nestin with purified ab176571 at 1:1000 dilution (1.65 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
-
Immunocytochemistry/ Immunofluorescence analysis of U-87 MG (Human glioblastoma-astrocytoma epithelial cell) cells labeling Nestin with purified ab176571 at 1:150 dilution (10 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
-
ICC/IF image of ab176571 stained SK-N-SH (Human neuroblastoma cell line) cells.
The cells were fixed in 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab176571 at 5 µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti-rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
-
Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling Nestin using ab176571 at a 1/250 dilution.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
-
Intracellular Flow Cytometry analysis of SH-SY5Y (human neuroblastoma) cells labeling Nestin with purified ab176571 at 1/160 dilution (10µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
-
Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Nestin using ab176571 at a 1/250 dilution.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (6)
ab176571 has been referenced in 6 publications.
- Notaras M et al. Schizophrenia is defined by cell-specific neuropathology and multiple neurodevelopmental mechanisms in patient-derived cerebral organoids. Mol Psychiatry 27:1416-1434 (2022). PubMed: 34789849
- Chen RH et al. Dedifferentiation of human epidermal melanocytes in vitro by long-term trypsinization. Cell Tissue Bank 22:67-75 (2021). PubMed: 32978700
- Meneceur S et al. Establishment and Characterisation of Heterotopic Patient-Derived Xenografts for Glioblastoma. Cancers (Basel) 12:N/A (2020). PubMed: 32260145
- Cohen MA et al. Formation of Human Neuroblastoma in Mouse-Human Neural Crest Chimeras. Cell Stem Cell 26:579-592.e6 (2020). PubMed: 32142683
- Wang J et al. Glioblastoma extracellular vesicles induce the tumour-promoting transformation of neural stem cells. Cancer Lett N/A:N/A (2019). PubMed: 31521694
- Chen C et al. Neuropathy correlated with imbalanced Foxp3/IL-17 in bone marrow microenvironment of patients with acute myeloid leukemia. Oncotarget 7:24455-65 (2016). IHC-P ; Human . PubMed: 27016413