Nestin Recombinant Rabbit Monoclonal Antibody [PSH07-89]
Catalog# HA722919
Nestin Recombinant Rabbit Monoclonal Antibody [PSH07-89]
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WB
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IF-Cell
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IHC-P
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IHC-Fr
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IF-Tissue
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mIHC
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Human
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Mouse
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Rat
Overview
Product Name
Nestin Recombinant Rabbit Monoclonal Antibody [PSH07-89]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within mouse Nestin aa 700-1,000.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, IHC-Fr, IF-Tissue, mIHC
Molecular Weight
Predicted band size: 207 kDa
Positive Control
RD cell lysate, Neuro-2a cell lysate, Mouse brain (P0) tissue lysate, Mouse brain (P7) tissue lysate, Mouse brain tissue lysate, U-87 MG, C2C12, C6, human kidney tissue, mouse kidney tissue, rat kidney tissue, mouse embryonic brain tissue.
Conjugation
unconjugated
Clone Number
PSH07-89
Product Features
Form
Liquid
Concentration
1ug/ul
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
-
1:2,000
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IF-Cell
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1:1,000-1:5,000
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IHC-P
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1:1,000
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IHC-Fr
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1:500-1:2,000
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IF-Tissue
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1:500
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mIHC
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1:1,000
Target
Function
Nestin is a major intermediate filament (IF) protein of embryonic central nervous system progenitor cells. It is also a component of the dynamic IF network during muscle development, where it polymerizes with Desmin and Vimentin. Nestin co-assembles with Vimentin or a-internexin and forms heterodimer coiled-coil molecules which then further assemble into 10 nml IFs. Deletion of the IF consensus rod domain in nestin alters nestin localization in CNS precursor cells and radial glial cells in vivo. Nestin is a marker for neuroepithelial stem cells, glioma cells and tumor endothelial cells during rapid growth. During axon elongation of differen-tiation neurons, nestin localizes to the growth cones and may play a role in growth cone guidance. In the rat adrenal gland, nestin is expressed by the zona fasciculata and the zona reticularis. Nestin is also expressed by dermatomal cells and by myoblasts during the earliest stages of myogenesis.
Background References
1. Yang Y et al. Heightened potency of human pluripotent stem cell lines created by transient BMP4 exposure. Proc Natl Acad Sci U S A 112:E2337-46 (2015).
2. Chen G et al. Human Brat ortholog TRIM3 is a tumor suppressor that regulates asymmetric cell division in glioblastoma. Cancer Res 74:4536-48 (2014).
Subcellular Location
Cytoplasm, Intermediate filament.
Synonyms
ESTM 46 antibody
FLJ 21841 antibody
FLJ21841 antibody
Intermediate filament protein antibody
Nbla00170 antibody
nes antibody
NEST_HUMAN antibody
Nestin antibody
Images
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Application: IHC-Fr
Species: Mouse
Site: E14.5 embryonic brain
Sample: Frozen section
Antibody concentration: 1/500
Antigen retrieval: Not required -
Application: IHC-Fr
Species: Rat
Site: Kindey
Sample: Frozen section
Antibody concentration: 1/500
Antigen retrieval: Not required -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Nestin antibody (HA722919) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722919) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Nestin antibody (HA722919) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722919) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Nestin antibody (HA722919) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722919) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse embryonic brain tissue with Rabbit anti-Nestin antibody (HA722919) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722919) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Application: IF-tissue
Species: Mouse
Site: E14.5 embryonic brain
Sample: Paraffin-embedded section
Antibody concentration: 1/500 -
☑ Relative expression (RE)
Western blot analysis of Nestin on different lysates with Rabbit anti-Nestin antibody (HA722919) at 1/2,000 dilution.
Lane 1: RD cell lysate
Lane 2: HEK-293 cell lysate (negative)
Lane 3: Neuro-2a cell lysate
Lane 4: Mouse brain (P0) tissue lysate
Lane 5: Mouse brain (P7) tissue lysate
Lane 6: Mouse brain tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 177/207 kDa
Observed band size: 150-300 kDa
Exposure time: 15 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722919) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of Nestin on different lysates with Rabbit anti-Nestin antibody (HA722919) at 1/1,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Nestin KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 177 kDa
Observed band size: 150-300 kDa
Exposure time: 20 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722919) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of C2C12 cells labeling Nestin with Rabbit anti-Nestin antibody (HA722919) at 1/5,000 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Nestin antibody (HA722919) at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C6 cells labeling Nestin with Rabbit anti-Nestin antibody (HA722919) at 1/1,000 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Nestin antibody (HA722919) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Products with the same target and pathway
Nestin Recombinant Rabbit Monoclonal Antibody [SN06-27]
Application: IF-Cell,IF-Tissue,IHC-P,WB
Reactivity: Human
Conjugate: unconjugated
Nestin Recombinant Antibody [PSH07-89] - Rat IgG1 (Chimeric)
Application: IHC-Fr,IHC-P,WB,IF-Cell
Reactivity: Human,Mouse,Rat
Conjugate: unconjugated
