Recombinant
RabMAb

Recombinant Anti-Myoferlin antibody [EPR18887] (ab178386)

Overview

  • Product name

    Anti-Myoferlin antibody [EPR18887]
    See all Myoferlin primary antibodies
  • Description

    Rabbit monoclonal [EPR18887] to Myoferlin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Myoferlin aa 1-100. The exact sequence is proprietary.
    Database link: Q9NZM1

  • Positive control

    • WB: MCF7, MDA-MB-231, HeLa, C6, NIH/3T3 and C2C12 whole cell lysates; Human fetal kidney and fetal heart lysates; Rat kidney lysate. ICC/IF: HeLa and C2C12 cells. Flow Cyt: HeLa cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18887
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab178386 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
WB 1/2000. Detects a band of approximately 250,180 kDa (predicted molecular weight: 235 kDa).
ICC/IF 1/100.
Flow Cyt 1/400.

Target

  • Function

    Calcium/phospholipid-binding protein that plays a role in the plasmalemma repair mechanism of endothelial cells that permits rapid resealing of membranes disrupted by mechanical stress. Involved in endocytic recycling. Implicated in VEGF signal transduction by regulating the levels of the receptor KDR.
  • Tissue specificity

    Expressed in myoblast and endothelial cells (at protein level). Highly expressed in cardiac and skeletal muscles. Also present in lung, and at very low levels in kidney, placenta and brain.
  • Sequence similarities

    Belongs to the ferlin family.
    Contains 5 C2 domains.
  • Domain

    The C2 domain 1 associates with lipid membranes in a calcium-dependent manner.
  • Cellular localization

    Cell membrane. Nucleus membrane. Cytoplasmic vesicle membrane. Concentrated at the membrane sites of both myoblast-myoblast and myoblast-myotube fusions. Detected at the plasmalemma in endothelial cells lining intact blood vessels (By similarity). Found at nuclear and plasma membranes. Enriched in undifferentiated myoblasts near the plasma membrane in puncate structures.
  • Information by UniProt
  • Database links

  • Alternative names

    • Fer 1 like 3, myoferlin (C. elegans) antibody
    • Fer 1 like family member 3 antibody
    • Fer 1 like protein 3 antibody
    • Fer-1-like protein 3 antibody
    • FER1L3 antibody
    • myoF antibody
    • MYOF_HUMAN antibody
    • Myoferlin antibody
    see all

Images

  • Lanes 1-2 : Anti-Myoferlin antibody [EPR18887] (ab178386) at 1/20000 dilution
    Lanes 3-5 : Anti-Myoferlin antibody [EPR18887] (ab178386) at 1/2000 dilution

    Lane 1 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
    Lane 2 : MDA-MB-231 (human breast adenocarcinoma cell line) whole cell lysate
    Lane 3 : Human fetal kidney lysate
    Lane 4 : Human fetal heart lysate
    Lane 5 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lanes 4-5 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

    Predicted band size: 235 kDa
    Observed band size: 180,250 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1/5: 10 seconds; Lane 2: 15 seconds; Lane 3/4: 3 minutes.

    The molecular weight  observed is consistent with what has been described in the literature (PMID: 22135466, PMID: 23499551).

  • Immunofluorescent analysis of 100% methanol-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Myoferlin with ab178386 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab178386 at 1/100 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.

    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Myoferlin with ab178386 at 1/400 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • All lanes : Anti-Myoferlin antibody [EPR18887] (ab178386) at 1/2000 dilution

    Lane 1 : Rat kidney lysate
    Lane 2 : C6 (rat glial tumor cell line) whole cell lysate
    Lane 3 : NIH/3T3 (mouse embryonic fibroblast cell line) whole cell lysate
    Lane 4 : C2C12 (mouse myoblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 235 kDa
    Observed band size: 180,250 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1:3 minutes; Lane 2/4: 10 seconds; Lane 3: 3 seconds.

     

  • Immunofluorescent analysis of 100% methanol-fixed C2C12 (mouse myoblast cell line) cells labeling Myoferlin with ab178386 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on C2C12 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab178386 at 1/100 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.

    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

References

ab178386 has not yet been referenced specifically in any publications.

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