Nestin Recombinant Rabbit Monoclonal Antibody [PSH07-89]
概述
产品名称
Nestin Recombinant Rabbit Monoclonal Antibody [PSH07-89]
抗体类型
Recombinant Rabbit monoclonal Antibody
免疫原
Recombinant protein within mouse Nestin aa 700-1,000.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IHC-P, IHC-Fr, IF-Tissue, mIHC, IP
分子量
Predicted band size: 207 kDa
阳性对照
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.
偶联
unconjugated
克隆号
PSH07-89
反应性数据
| WB | IF-Cell | IHC-P | IHC-Fr | IF-Tissue | mIHC | IP | |
|---|---|---|---|---|---|---|---|
| Human |
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| Mouse |
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| Rat |
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产品特性
形态
Liquid
存放说明
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
存储缓冲液
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
亚型
IgG
纯化方式
Protein A affinity purified.
应用稀释度
-
WB
-
1:2,000
-
IF-Cell
-
1:1,000-1:5,000
-
IHC-P
-
1:1,000
-
IHC-Fr
-
1:500-1:2,000
-
IF-Tissue
-
1:500
-
mIHC
-
1:1,000
-
IP
-
Use at an assay dependent concentration.
靶点
功能
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.
背景文献
1. Chen H et al. Targeting Nestin(+) hepatic stellate cells ameliorates liver fibrosis by facilitating TbetaRI degradation. J Hepatol. 2021 May
2. Wang J et al. Nestin promotes pulmonary fibrosis via facilitating recycling of TGF-beta receptor I. Eur Respir J. 2022 May
亚细胞定位
Cytoplasm, Intermediate filament.
别名
ESTM 46 antibody
FLJ 21841 antibody
FLJ21841 antibody
Intermediate filament protein antibody
Nbla00170 antibody
nes antibody
NEST_HUMAN antibody
Nestin antibody
图片
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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 -
Application: IF-tissue
Species: Mouse
Site: E14.5 embryonic brain
Sample: Paraffin-embedded section
Antibody concentration: 1/500 -
Application: IF-tissue
Species: Mouse
Site: embryos (cartilage)
Sample: Paraffin-embedded section
Antibody concentration: 1/500 -
Application: IF-tissue
Species: Rat
Site: brain
Sample: Paraffin-embedded section
Antibody concentration: 1/500 -
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. -
☑ 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. -
Nestin was immunoprecipitated from 0.2 mg C2C12 cell lysate with HA722919 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722919 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: C2C12 cell lysate (input)
Lane 2: HA722919 IP in C2C12 cell lysate
Lane 3: Rabbit IgG instead of HA722919 in C2C12 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 6 seconds; ECL: K1801
请注意: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
引文
-
Multimerin-1 modulates macrophage M2 polarization and enhances tumor cell stemness in glioblastoma
期刊: Neurological Research
DOI: 10.1080/01616412.2025.2520060
IF: 1.5
应用: WB
反应种属: Human
发表时间: 2025 Jun
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