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遺伝子ノックアウト細胞株 検証済

Anti-Sortilin/NT3 抗体 (ab16640)

  • Datasheet
  • SDS
Reviews (9)Q&A (2)References (79)

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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

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-Sortilin/NT3 antibody
    Sortilin/NT3 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to Sortilin/NT3
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: IHC-FoFr, WBmore details
  • 種交差性

    交差種: Mouse, Rat, Human
    交差が予測される動物種: Cow
  • 免疫原

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

  • ポジティブ・コントロール

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

    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

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    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.
  • バッファー

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    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...
  • 精製度

    Immunogen affinity purified
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipoproteins/Apolipoproteins
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    • Heart disease

関連製品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • 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)
  • KO cell pellets

    • Human SORT1 (Sortilin/NT3) knockout HeLa cell pellet (ab278985)
  • Recombinant Protein

    • Recombinant Rat Sortilin/NT3 protein (His tag) (ab226427)

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab16640の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
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).
特記事項
IHC-FoFr
1/3000.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 95 kDa).

ターゲット情報

  • 機能

    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.
  • 組織特異性

    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.
  • 関連疾患

    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.
  • 配列類似性

    Belongs to the VPS10-related sortilin family. SORT1 subfamily.
    Contains 9 BNR repeats.
  • ドメイン

    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.
  • 翻訳後修飾

    The N-terminal propeptide is cleaved by furin and possibly other homologous proteases.
  • 細胞内局在

    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
  • 参照データベース

    • 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
  • 別名

    • 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

画像

  • 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

プロトコール

  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

データシートおよび資料

  • SDS download

  • Datasheet download

    Download

参考文献 (79)

ab16640 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab16640 は 79 報の論文で使用されています。

  • van Erp S  et al. Age-related loss of axonal regeneration is reflected by the level of local translation. Exp Neurol 339:113594 (2021). PubMed: 33450233
  • Berger K  et al. Tumor co-expression of progranulin and sortilin as a prognostic biomarker in breast cancer. BMC Cancer 21:185 (2021). PubMed: 33618683
  • Fleury S  et al. Tissue-Specificity of Antibodies Raised Against TrkB and p75NTR Receptors; Implications for Platelets as Models of Neurodegenerative Diseases. Front Immunol 12:606861 (2021). PubMed: 33643311
  • Liang M  et al. Circular RNA hsa_circ_0110389 promotes gastric cancer progression through upregulating SORT1 via sponging miR-127-5p and miR-136-5p. Cell Death Dis 12:639 (2021). PubMed: 34162830
  • Yasa S  et al. CLN3 regulates endosomal function by modulating Rab7A-effector interactions. J Cell Sci 133:N/A (2020). PubMed: 32034082
View all Publications for this product

レビューと Q&A

Show All レビュー Q&A
レビューを送る 質問を送る

1-10 of 11 Abreviews or Q&A

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

Abcam user community

Verified customer

投稿 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

Abcam user community

Verified customer

投稿 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

Abcam user community

Verified customer

投稿 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

Abcam user community

Verified customer

投稿 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

DR. Ruma Raha-Chowdhury

Verified customer

投稿 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

DR. Ruma Raha-Chowdhury

Verified customer

投稿 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

Mr. Youngjeon Lee

Verified customer

投稿 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

Matthew Provenzano

Verified customer

投稿 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

Answered on Oct 05 2012

Question

I am still quite concerned/convinced that the anti-sortilin (16640) antibody is not really an antibody to sortilin. (Images attached). Your suggestion of "increasing the incubation time from 30 min to overnight to see if the signal is getting stronger and more specific" results in the expected massive label which again appears to be cytoskeletal with no specificity for Golgi or endocytic compartments. I have also compared the ab16640 to another commercially available anti-Sortilin antibody (this antibody labels the expected sortilin-positive compartments by IFA and recognizes the appropriate band by Western.) In the future if you discover that the lot of antibody I received was incorrect (104822), please keep me in mind for a replacement. In addition, your ab16640 has very interesting localization in bacterial infected cells. It may be cross-reacting with bacteria, but if not, I would love to know what this antibody recognizes if it is other than sortilin.

Read More

Abcam community

Verified customer

Asked on Nov 01 2006

Answer

Thank you for getting back to me and for sharing your recent findings with me. I am very sorry to hear that even after following my suggestions, you are still not satisfied with this antibody. This product has been tested for IHC by one of our collaborators who has used rat cortical neurons fixed with 4% paraformaldehyde. Rats were intracardially perfused with 4% paraformaldehyde and the 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 performed in free floating with fixed tissues (rat brain sections). As I understand from your previous e-mail, you tried 1% formaldehyde, 1% paraformaldehyde, 4% paraformaldehyde for 20 minutes. As I have mentioned ab16640 has not been tested for ICC/IF, only for IHC on perfused samples and it is very likely that the different fixation technique may be the reason for this discrepancy. My last suggestion, perhaps, would be to test ice-cold methanol or acetone (only for 5 mins) as alternative fixation to see if it changes the staining pattern. Since you purchased this antibody in September and the guarantee period (as Abcam promise indicates) has not expired, I am ready to offer you a new vial as a free of charge replacement – in case the product may have been damaged during storing or shipping. Please do let me know if you would like to get one vial of this product to test or not. Your findings are very interesting and I am convinced that other customers would find your feedback very useful. We always encourage customers to send their results back to us, whether positive or negative, and make all product information available to researchers. If you send us an Abreview, we can offer you from 50 up to 300 Abcam points (Abpoints) based on submission time and type of application. The less characterised the product you review, the more Abpoints you will receive. Furthermore, you get 100 extra Abpoints for submitting an image. These Abpoints can then be claimed back as rewards. To find out more about our new Loyalty Scheme, Abpoints and rewards please visit our website at www.abcam.com/abreviews and look at Your Account panel (left bottom section on our home page). You can submit the Abreview if you click on the Abreview section (top menu bar) on the on-line datasheet of this antibody. It is a very straightforward process. Should you need any help with your submission, then please do not hesitate to contact us again. Thank you for contacting Abcam again and I am looking forward to hearing from you.

Read More

Abcam Scientific Support

Answered on Nov 03 2006

1-10 of 11 Abreviews or Q&A

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  • 癌
  • 循環器
  • 細胞生物学
  • 発生
  • エピジェネティクス&核内情報
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