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    products/primary-antibodies/furin-antibody-ab3467.pdf

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Signal Transduction Metabolism Amino Acids
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Anti-Furin antibody (ab3467)

  • Datasheet
Reviews (7)Q&A (2)References (21)

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Immunocytochemistry - Anti-Furin antibody (ab3467)
  • Western blot - Anti-Furin antibody (ab3467)
  • Immunocytochemistry - Anti-Furin antibody (ab3467)
  • Western blot - Anti-Furin antibody (ab3467)
  • Immunocytochemistry - Anti-Furin antibody (ab3467)

Key features and details

  • Rabbit polyclonal to Furin
  • Suitable for: ICC, WB
  • Reacts with: Mouse, Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Furin antibody
    See all Furin primary antibodies
  • Description

    Rabbit polyclonal to Furin
  • Host species

    Rabbit
  • Specificity

    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.
  • Tested applications

    Suitable for: ICC, WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    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
  • General notes

    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

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

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

    Immunogen affinity purified
  • Primary antibody notes

    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.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Amino Acids
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cell Adhesion
    • Integrins
    • Alpha
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteolytic enzymes
    • Serine protease
    • Proprotein Convertases

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunizing Peptide (Blocking)

    • Furin peptide (ab4989)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • Recombinant Furin protein (ab6032)
  • Recombinant Protein

    • Recombinant Furin protein (ab6032)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
ICC
Use a concentration of 2 µg/ml.
WB (3)
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.

Notes
ICC
Use a concentration of 2 µg/ml.
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.

Target

  • Function

    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.
  • Tissue specificity

    Seems to be expressed ubiquitously.
  • Sequence similarities

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

    Contains a cytoplasmic domain responsible for its TGN localization and recycling from the cell surface.
  • Post-translational
    modifications

    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.
  • Cellular localization

    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.
  • Target information above from: UniProt accession P09958 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5045 Human
    • Entrez Gene: 18550 Mouse
    • Omim: 136950 Human
    • SwissProt: P09958 Human
    • SwissProt: P23188 Mouse
    • Unigene: 513153 Human
    • Unigene: 5241 Mouse
    • Alternative names

      • 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

    Images

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

      Immunocytochemical analysis of methanol-fixed 2% BSA blocked HeLa cells strining furin with ab3467 at 2 µg/ml and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution (green). Nuclei are stained with SlowFade® Gold Antifade Mountant with DAPI (blue), and the cytoskeleton with alpha-Tubulin Monoclonal Antibody and Goat anti-Mouse IgG Secondary Antibody, Alexa Fluor® 594 conjugate at 1/400 dilution (red). Negative control uses no primary antibody

    • Western blot - Anti-Furin antibody (ab3467)
      Western blot - Anti-Furin antibody (ab3467)
      All lanes : Anti-Furin antibody (ab3467) at 1/500 dilution

      Lane 1 : HeLa lysate at 30 µg
      Lane 2 : HEK-293 extract at 30 µg
      Lane 3 : K-562 extract at 30 µg
      Lane 4 : SH-SY5Y extract at 30 µg
      Lane 5 : conditioned media from HeLa cell line at 10 µg

      Secondary
      All lanes : Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 1/2500 dilution

      Developed using the ECL technique.

      Predicted band size: 87 kDa

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

    Protocols

    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (21)

    Publishing research using ab3467? Please let us know so that we can cite the reference in this datasheet.

    ab3467 has been referenced in 21 publications.

    • O'Sullivan MJ  et al. In well-differentiated primary human bronchial epithelial cells, TGF-ß1 and TGF-ß2 induce expression of furin. Am J Physiol Lung Cell Mol Physiol 320:L246-L253 (2021). PubMed: 33174447
    • Papa G  et al. Furin cleavage of SARS-CoV-2 Spike promotes but is not essential for infection and cell-cell fusion. PLoS Pathog 17:e1009246 (2021). PubMed: 33493182
    • Wang H  et al. The role of high cholesterol in age-related COVID19 lethality. bioRxiv N/A:N/A (2021). PubMed: 32511366
    • Akerman AW  et al. miR-133a Replacement Attenuates Thoracic Aortic Aneurysm in Mice. J Am Heart Assoc 10:e019862 (2021). PubMed: 34387094
    • Ueo A  et al. Lysosome-Associated Membrane Proteins Support the Furin-Mediated Processing of the Mumps Virus Fusion Protein. J Virol 94:N/A (2020). PubMed: 32295904
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-Furin antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hek293T)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Hek293T
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°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

    Submitted Jul 09 2020

    Western blot abreview for Anti-Furin antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    African green monkey Cell lysate - other (Vero)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Vero
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°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

    Submitted Jul 08 2020

    Western blot abreview for Anti-Furin antibody

    Excellent
    Abreviews
    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
    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. Lawrence Welch

    Verified customer

    Submitted May 07 2020

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