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使用敲除细胞株进行验证

Anti-Sortilin/NT3抗体(ab16640)

  • Datasheet
  • SDS
Reviews (10)Q&A (2)References (94)

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Western blot - Anti-Sortilin/NT3 antibody (ab16640)
  • Western blot - Anti-Sortilin/NT3 antibody (ab16640)
  • Western blot - Anti-Sortilin/NT3 antibody (ab16640)
  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Sortilin/NT3 antibody (ab16640)

Key features and details

  • Rabbit polyclonal to Sortilin/NT3
  • Suitable for: IHC-FoFr, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

选择批间可重复性更高的重组抗体

Product image
Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-Sortilin/NT3抗体
    参阅全部 Sortilin/NT3 一抗
  • 描述

    兔多克隆抗体to Sortilin/NT3
  • 宿主

    Rabbit
  • 经测试应用

    适用于: IHC-FoFr, WBmore details
  • 种属反应性

    与反应: Mouse, Rat, Human
    预测可用于: Cow
  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: HeLa cell lysate; NIH/3T3 cell lysate; 293T cell lysate; Human, Mouse, and Rat Brain tissue lysates. IHC-FoFr: Rat cortical neurons.
  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 98.98% PBS, 1% BSA

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Cardiovascular
    • Lipids / Lipoproteins
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相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • KO cell lines

    • Human SORT1 (Sortilin/NT3) knockout HeLa cell line (ab264772)
  • KO cell lysates

    • Human SORT1 (Sortilin/NT3) knockout HeLa cell lysate (ab257696)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab16640于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
IHC-FoFr (2)
1/3000.
WB (5)
Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 95 kDa).
说明
IHC-FoFr
1/3000.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 95 kDa).

靶标

  • 功能

    Functions as a sorting receptor in the Golgi compartment and as a clearance receptor on the cell surface. Required for protein transport from the Golgi apparatus to the lysosomes by a pathway that is independent of the mannose-6-phosphate receptor (M6PR). Also required for protein transport from the Golgi apparatus to the endosomes. Promotes neuronal apoptosis by mediating endocytosis of the proapoptotic precursor forms of BDNF (proBDNF) and NGFB (proNGFB). Also acts as a receptor for neurotensin. May promote mineralization of the extracellular matrix during osteogenic differentiation by scavenging extracellular LPL. Probably required in adipocytes for the formation of specialized storage vesicles containing the glucose transporter SLC2A4/GLUT4 (GLUT4 storage vesicles, or GSVs). These vesicles provide a stable pool of SLC2A4 and confer increased responsiveness to insulin. May also mediate transport from the endoplasmic reticulum to the Golgi.
  • 组织特异性

    Expressed at high levels in brain, spinal cord, heart, skeletal muscle, thyroid, placenta and testis. Expressed at lower levels in lymphoid organs, kidney, colon and liver.
  • 疾病相关

    Note=A common polymorphism located in a non-coding region between CELSR2 and PSRC1 alters a CEBP transcription factor binding site and is responsible for changes in hepatic expression of SORT1. Altered SORT1 expression in liver affects low density lipoprotein cholesterol levels in plasma and is associated with susceptibility to myocardial infarction.
  • 序列相似性

    Belongs to the VPS10-related sortilin family. SORT1 subfamily.
    Contains 9 BNR repeats.
  • 结构域

    The N-terminal propeptide may facilitate precursor transport within the Golgi stack. Intrachain binding of the N-terminal propeptide and the extracellular domain may also inhibit premature ligand binding.
    The extracellular domain may be shed following protease cleavage in some cell types.
  • 翻译后修饰

    The N-terminal propeptide is cleaved by furin and possibly other homologous proteases.
  • 细胞定位

    Membrane. Endoplasmic reticulum membrane. Endosome membrane. Golgi apparatus > Golgi stack membrane. Lysosome membrane. Nucleus membrane. Cell membrane. Lysosome membrane. Localized to membranes of the endoplasmic reticulum, endosomes, Golgi stack, lysosomes and nucleus. A small fraction of the protein is also localized to the plasma membrane. May also be found in SLC2A4/GLUT4 storage vesicles (GSVs) in adipocytes. Localization to the plasma membrane in adipocytes may be enhanced by insulin.
  • Target information above from: UniProt accession Q99523 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 6272 Human
    • Entrez Gene: 20661 Mouse
    • Entrez Gene: 83576 Rat
    • Omim: 602458 Human
    • SwissProt: Q99523 Human
    • SwissProt: Q6PHU5 Mouse
    • SwissProt: O54861 Rat
    • Unigene: 485195 Human
    • Unigene: 703487 Human
    • Unigene: 157119 Mouse
    • Unigene: 11286 Rat
    see all
  • 别名

    • 100 kDa NT receptor antibody
    • Glycoprotein 95 antibody
    • Gp 95 antibody
    • Gp95 antibody
    • LDLCQ6 antibody
    • Neurotensin receptor 3 antibody
    • NT 3 antibody
    • NT3 antibody
    • NTR 3 antibody
    • NTR3 antibody
    • OTTHUMP00000013784 antibody
    • SORT 1 antibody
    • SORT_HUMAN antibody
    • SORT1 (gene name) antibody
    • Sort1 antibody
    • Sortilin 1 antibody
    • Sortilin antibody
    see all

图片

  • Western blot - Anti-Sortilin/NT3 antibody (ab16640)
    Western blot - Anti-Sortilin/NT3 antibody (ab16640)
    All lanes : Anti-Sortilin/NT3 antibody (ab16640) at 1 µg/ml

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : SORT1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 95 kDa
    Observed band size: 100 kDa why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab16640 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab16640 was shown to react with Sortilin/NT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264772 (knockout cell lysate ab257696) was used. Wild-type HeLa and SORT1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab16640 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Sortilin/NT3 antibody (ab16640)
    Western blot - Anti-Sortilin/NT3 antibody (ab16640)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Sortilin/NT3 knockout HAP1 cell lysate (20 µg)
    Lane 3: NIH/3T3 cell lysate (20 µg)
    Lane 4: 293T cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab16440 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.


    ab16640 was shown to specifically react with Sortilin/NT3 in wild-type HAP1 cells. No band was observed when Sortilin/NT3 knockout samples were examined. Wild-type and Sortilin/NT3 knockout samples were subjected to SDS-PAGE. ab16640 and ab8245 (loading control to GAPDH) were diluted 1 µg/mL and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Sortilin/NT3 antibody (ab16640)
    Western blot - Anti-Sortilin/NT3 antibody (ab16640)
    Lane 1 : Marker
    Lanes 2-4 : Anti-Sortilin/NT3 antibody (ab16640) at 1 µg/ml

    Lane 2 : Human Brain tissue lysate at 20 µg
    Lane 3 : Mouse Brain tissue lysate at 20 µg
    Lane 4 : Rat Brain whole cell lysate at 20 µg

    Secondary
    Lanes 2-4 : Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 95 kDa
    Additional bands at: 31 kDa (possible cleavage fragment)

  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Sortilin/NT3 antibody (ab16640)
    Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Sortilin/NT3 antibody (ab16640)This image is courtesy of Sophie Pezet, Univ London Kings Coll, United Kingdom

    Immunofluorescent staining for Sortilin/NT3 in rat cortical neurons using ab16640. The staining is cytoplasmic and punctate of many cells such as cortical neurons. The image was taken with a X20 objective.

    Protocol details: Rats were intracardially perfused with 4% paraformaldehyde. Whole brain tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT and then cut on cryostat (30µm coronal sections). IHC was perfored in free floating with fixed tissues (rat brain sections). Primary antibody was incubated overnight at 1/3000 at room temperature

实验方案

  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (94)

发表研究结果有使用 ab16640?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab16640 被引用在 94 文献中.

  • Tran ML  et al. Cab45 deficiency leads to the mistargeting of progranulin and prosaposin and aberrant lysosomal positioning. Traffic 24:4-19 (2023). PubMed: 36398980
  • Jing Y  et al. Spinal cord injury-induced gut dysbiosis influences neurological recovery partly through short-chain fatty acids. NPJ Biofilms Microbiomes 9:99 (2023). PubMed: 38092763
  • Lui A  et al. Regulation of Human Sortilin Alternative Splicing by Glucagon-like Peptide-1 (GLP1) in Adipocytes. Int J Mol Sci 24:N/A (2023). PubMed: 37762628
  • Jing Y  et al. Fecal Microbiota Transplantation Exerts Neuroprotective Effects in a Mouse Spinal Cord Injury Model by Modulating the Microenvironment at the Lesion Site. Microbiol Spectr 10:e0017722 (2022). PubMed: 35467388
  • Yamamoto T  et al. SORT1/LAMP2-mediated extracellular vesicle secretion and cell adhesion are linked to lenalidomide resistance in multiple myeloma. Blood Adv 6:2480-2495 (2022). PubMed: 34979567
View all Publications for this product

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1-10 of 12 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Sortilin/NT3 antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (colon tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Target Retrieval Solution (TRS) pH 6
Permeabilization
No
Specification
colon tumor
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 2.5% · Temperature: 22°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Oct 14 2022

Western blot abreview for Anti-Sortilin/NT3 antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (kidney)
Gel Running Conditions
Reduced Denaturing (4-12% bis-tris gel)
Loading amount
30 µg
Specification
kidney
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Nov 03 2020

Western blot abreview for Anti-Sortilin/NT3 antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Dopaminergic Neurons)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Dopaminergic Neurons
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Oct 22 2020

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Sortilin/NT3 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric buffer
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 May 27 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-Sortilin/NT3 antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Astrocytes)
Permeabilization
No
Specification
Astrocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 37°C
Fixative
Methanol
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 May 26 2020

Western blot abreview for Anti-Sortilin/NT3 antibody

Below Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Tissue lysate - whole (Rat brain-Hippocampus, cortex)
Gel Running Conditions
Non-reduced Denaturing (4-12 % Bis-Tris)
Loading amount
20 µg
Specification
Rat brain-Hippocampus, cortex
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Ruma Raha-Chowdhury

Verified customer

提交于 Apr 30 2012

Immunocytochemistry abreview for Anti-Sortilin/NT3 antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Astrocytes culture)
Permeabilization
Yes - 0.3% TritonX in 0.1% PBS
Specification
Astrocytes culture
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Ruma Raha-Chowdhury

Verified customer

提交于 Feb 13 2012

Western blot abreview for Anti-Sortilin/NT3 antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
Brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Mr. Youngjeon Lee

Verified customer

提交于 Dec 29 2009

Western blot abreview for Anti-Sortilin antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Schwann Cells)
Loading amount
45 µg
Specification
Schwann Cells
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Matthew Provenzano

Verified customer

提交于 Mar 27 2007

Question

I see multiple bands in blots of various samples, including an over-expressed positive control. The band corresponding to the molecular weight of sortilin is relatively weak. Previous vials have performed much better.

Read More

Abcam community

Verified customer

Asked on Oct 05 2012

Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number ****.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Abcam Scientific Support

回复于 Oct 05 2012

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