Anti-Furin 抗体 (ab3467)

Reviews (3)Q&A (2)References (14)

製品の概要

  • 製品名

    Anti-Furin antibody
    Furin 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Furin

  • 由来種

    Rabbit

  • 特異性

    Detects Furin from canine and mouse cells as well as transfected human Furin. This antibody does not detect endogenous Furin from BSC-40, HeLa, J774A.1 BPAEC, or CHO cells nor from rat skeletal muscle, spleen, kidney, ovary, testes, heart, or brain tissue.

  • アプリケーション

    適用あり: ICC, Inhibition Assay, ICC/IF, IP, WB, IHC-Pmore details

  • 種交差性

    交差種: Mouse, Rat, Sheep, Hamster, Dog, Human, Pig, Non human primates

  • 免疫原

    Synthetic peptide corresponding to Human Furin aa 780-793.
    Sequence:

    RGERTAFIKDQSAL


    (Peptide available as ab4989)

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

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

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...

  • 精製度

    Immunogen affinity purified

  • 一次抗体 備考

    Furin is a membrane-associated, calcium-dependent, serine protease that belongs to the subtilisin-like prohormone convertase (PC) family. Members of this family of cellular enzymes cleave most prohormones and neuropeptide precursors. Numerous other cellular proteins, some viral proteins, and bacterial toxins that are transported by the constitutive secretory pathway are also targeted for maturation by PCs. Furin and other PC family members share structural similarities which include a heterogeneous ~10 kDa amino-terminal proregion, a highly conserved ~55 kDa subtilisin-like catalytic domain, and carboxyl-terminal domain that is heterogeneous in length and sequence. These enzymes become catalytically active following proregion cleavage within the appropriate cellular compartment. Furin is the only known PC to possess a transmembrane domain. Cleavage of target proteins occurs at the carboxyl-terminus of the furin consensus sequence, RX(K/R)R. It has been shown that the acidic peptide sequence, C771PSDSEEDEG780, localizes furin to the trans-Golgi-network. Phosphorylation of serine residues within this region modulates intracellular routing of Furin protein. An additional signaling domain includes the tetrapeptide sequence, Y759KGL762, which directs internalization from the cell surface.

  • ポリ/モノ

    ポリクローナル

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab3467 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC Use at an assay dependent concentration.
Inhibition Assay Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 18174282
IP 1/100.
WB Use at an assay dependent concentration. Detects a band of approximately 100 kDa (predicted molecular weight: 87 kDa).Can be blocked with Furin peptide (ab4989).

Representing Furin in BSC-40 cells transfected with the human Furin gene.
IHC-P Use at an assay dependent concentration. PubMed: 23418414

ターゲット情報

  • 機能

    Furin is likely to represent the ubiquitous endoprotease activity within constitutive secretory pathways and capable of cleavage at the RX(K/R)R consensus motif.

  • 組織特異性

    Seems to be expressed ubiquitously.

  • 配列類似性

    Belongs to the peptidase S8 family. Furin subfamily.
    Contains 1 homo B/P domain.

  • ドメイン

    Contains a cytoplasmic domain responsible for its TGN localization and recycling from the cell surface.

  • 翻訳後修飾

    The inhibition peptide, which plays the role of an intramolecular chaperone, is autocatalytically removed in the endoplasmic reticulum (ER) and remains non-covalently bound to furin as a potent autoinhibitor. Following transport to the trans Golgi, a second cleavage within the inhibition propeptide results in propeptide dissociation and furin activation.
    Phosphorylation is required for TGN localization of the endoprotease. In vivo, exists as di-, mono- and non-phosphorylated forms.

  • 細胞内局在

    Golgi apparatus > trans-Golgi network membrane. Cell membrane. Shuttles between the trans-Golgi network and the cell surface. Propeptide cleavage is a prerequisite for exit of furin molecules out of the endoplasmic reticulum (ER). A second cleavage within the propeptide occurs in the trans Golgi network (TGN), followed by the release of the propeptide and the activation of furin.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • Dibasic processing enzyme antibody
    • Dibasic-processing enzyme antibody
    • FES upstream region antibody
    • FUR antibody
    • FURIN antibody
    • Furin membrane associated receptor protein antibody
    • FURIN_HUMAN antibody
    • PACE antibody
    • Paired basic amino acid residue cleaving enzyme antibody
    • Paired basic amino acid residue-cleaving enzyme antibody
    • PCSK3 antibody
    • Proprotein convertase subtilisin/kexin type 3 antibody
    • SPC1 antibody
    see all

画像

  • Immunocytochemistry/ Immunofluorescence - Anti-Furin antibody (ab3467)
    Immunocytochemistry/ Immunofluorescence - Anti-Furin antibody (ab3467)

    Immunolocalization of endogenous furin in mouse 3T3 cells with ab3467.

  • Western blot - Anti-Furin antibody (ab3467)
    Western blot - Anti-Furin antibody (ab3467)
    All lanes : Anti-Furin antibody (ab3467) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 87 kDa
    Observed band size: 63 kDa
    why is the actual band size different from the predicted?


    Exposure time: 90 seconds


    The band observed at 63 kDa could potentially be a cleaved form of Furin due to the presence of a 24 amino acid signal peptide and a 83 amino acid propeptide.
  • Immunocytochemistry/ Immunofluorescence - Anti-Furin antibody (ab3467)
    Immunocytochemistry/ Immunofluorescence - Anti-Furin antibody (ab3467)Image courtesy of an anonymous Abreview.
    ab3467 staining Furin in human A549 cells by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed with paraformaldehyde, permeabilized using 0.1% Triton X-100, blocked with 1% BSA/ 5%FBS/PBS for 1 hour at 25°C and then incubated with ab3467 at a 1/100 dilution for 1 hour at 25°C. The secondary used was an Alexa-Fluor 546 conjugated goat anti-rabbit polyclonal used at a 1/500 dilution.

    Cells were counterstained with DAPI (436nM, 5 mins) and Phalloidin-633 (1unit/coverslip) and mounted with prolong gold. No staining seen in secondary control and furin staining was punctate and throughout the cytoplasm.

    See Abreview

参考文献

ab3467 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。

ab3467 は 14 報の論文で使用されています。

  • Chang JC  et al. SOST/Sclerostin Improves Posttraumatic Osteoarthritis and Inhibits MMP2/3 Expression After Injury. J Bone Miner Res 33:1105-1113 (2018). PubMed: 29377313
  • Zhao G  et al. Influence of a Coronary Artery Disease-Associated Genetic Variant on FURIN Expression and Effect of Furin on Macrophage Behavior. Arterioscler Thromb Vasc Biol 38:1837-1844 (2018). PubMed: 29976768
  • Knab VM  et al. Acute Parathyroid Hormone Injection Increases C-Terminal but Not Intact Fibroblast Growth Factor 23 Levels. Endocrinology 158:1130-1139 (2017). PubMed: 28324013
  • Delestre-Delacour C  et al. Myosin 1b and F-actin are involved in the control of secretory granule biogenesis. Sci Rep 7:5172 (2017). PubMed: 28701771
  • Zhou Z  et al. The cAMP-responsive element binding protein (CREB) transcription factor regulates furin expression during human trophoblast syncytialization. Placenta 35:907-18 (2014). PubMed: 25175744
View all Publications for this product

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Western blot abreview for Anti-Furin antibody

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HAP1)
Gel Running Conditions
Non-reduced Denaturing (4-20% tris-glycine)
Loading amount
250000 cells
Specification
HAP1
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
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Mr. Lawrence Welch

Verified customer

投稿 May 07 2020

Question

So, you are saying, the 60kD band is the Signal Sequence (˜24AA)+ Propeptide (˜130AA) + catalytic domain (˜280)+ partial RGD domain.



Thank you,

Read More
Answer

Sorry for this misunderstanding.

We believe that the antibody recognizes the mature form of the protein. The pro-peptide is expected at 87kDa and once the signal and pro-peptide are cleaved off the protein, it migrates at 63 kDa.

I hope this is helpful and wish you a good weekend.

Read More

Immunocytochemistry/ Immunofluorescence abreview for Anti-Furin antibody

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (3T3 fibroblasts)
Specification
3T3 fibroblasts
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% TX-100
Blocking step
1% BSA/ 5%FBS/PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More

Abcam user community

Verified customer

投稿 May 12 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-Furin antibody

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (A549 lung epithelial cell line)
Specification
A549 lung epithelial cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% TX-100
Blocking step
1% BSA/ 5%FBS/PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More

Abcam user community

Verified customer

投稿 May 12 2012

Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 111548 DESCRIPTION OF THE PROBLEM Antibody recognises multiple bands - major ones at 250, 100, 75 (triplet) and also picks up 37 kDa MW marker! Peptide (ab4989) takes out 250, upper band of 75 kDa triplet and MW marker (!). SAMPLE Human cultured cardiac fibroblasts - whole cell homogenate PRIMARY ANTIBODY Abcam furin convertase (ab3467) 1:500 - 1:1000 DETECTION METHOD Pierce ECL POSITIVE AND NEGATIVE CONTROLS USED HT-1080 cell homogenate ANTIBODY STORAGE CONDITIONS -20C aliquoted SAMPLE PREPARATION Laemmli sample buffer + PMSF + Na vanadate. Boiled + 2-mercaptoethanol for 5 min. AMOUNT OF PROTEIN LOADED 50-100 ug per lane ELECTROPHORESIS/GEL CONDITIONS 10% gel. reducing. TRANSFER AND BLOCKING CONDITIONS Blocked in 5% Milk in TBS Tween SECONDARY ANTIBODY [a competitor] rabbit-HRP 1:2000 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Antibody dilution, addition of neutralising peptide ADDITIONAL NOTES Attached is a figure of blot with and without peptide.

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Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. We have received a favourable review of our Furin Convertase antibody (ab3467) albeit with acknowledgement of the detection of some non-specific bands. We often receive complaints relating to aberrant bands that can often be resolved with the use of 5% BSA as a blocking agent. We find it gives a cleaner more specific blot. I would therefore like to recommend its use on this occasion. I would also recommend performing an overnight incubation of the antibody, at a more dilute concentration e.g. 1:1500 at 4oC as again we often find that this increases the specificity of the blot. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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