Overview

  • Product nameAnti-ALIX antibody
    See all ALIX primary antibodies
  • Description
    Rabbit polyclonal to ALIX
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Chicken, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human ALIX aa 450-550 conjugated to Keyhole Limpet Haemocyanin (KLH).
    (Peptide available as ab88953, ab88953)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa, HepG2, SHSY5Y and HUVEC. This antibody gave a positive result in IHC in the following FFPE tissue: Human breast cancer.

Properties

Applications

Our Abpromise guarantee covers the use of ab76608 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 Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 96 kDa).
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • FunctionClass E VPS protein involved in concentration and sorting of cargo proteins of the multivesicular body (MVB) for incorporation into intralumenal vesicles (ILVs) that are generated by invagination and scission from the limiting membrane of the endosome. Binds to the phospholipid lysobisphosphatidic acid (LBPA) which is abundant in MVBs internal membranes. The MVB pathway appears to require the sequential function of ESCRT-O, -I,-II and -III complexes. The ESCRT machinery also functions in topologically equivalent membrane fission events, such as the terminal stages of cytokinesis and enveloped virus budding (HIV-1 and other lentiviruses). Appears to be an adapter for a subset of ESCRT-III proteins, such as CHMP4, to function at distinct membranes. Required for completion of cytokinesis. Involved in HIV-1 virus budding. Can replace TSG101 it its role of supporting HIV-1 release; this function implies the interaction with CHMP4B. May play a role in the regulation of both apoptosis and cell proliferation.
  • Sequence similaritiesContains 1 BRO1 domain.
  • Cellular localizationCytoplasm > cytosol. Melanosome. Cytoplasm > cytoskeleton > centrosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Colocalized with CEP55 in the midbody during cytokinesis. Colocalized with CEP55 at centrosomes of non-dividing cells.
  • Information by UniProt
  • Database links
  • Alternative names
    • AIP1 antibody
    • ALG 2 interacting protein 1 antibody
    • ALG-2-interacting protein 1 antibody
    • ALG2 interacting protein X antibody
    • Alix antibody
    • Apoptosis linked gene 2 interacting protein X antibody
    • Dopamine receptor interacting protein 4 antibody
    • DRIP4 antibody
    • Hp95 antibody
    • KIAA1375 antibody
    • MGC17003 antibody
    • PDC6I_HUMAN antibody
    • PDCD6 interacting protein antibody
    • PDCD6-interacting protein antibody
    • PDCD6IP antibody
    • Programmed cell death 6 interacting protein antibody
    • Programmed cell death 6-interacting protein antibody
    see all

Anti-ALIX antibody images

  • All lanes : Anti-ALIX antibody (ab76608) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 4 : HUVEC (Human Umbilical Vein Endothelial Cell) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 96 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 48 kDa,90 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes
    Programmed cell death 6-interacting protein (PDC6I) contains a number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight (100 kDa) than predicted (96 kDa).
  • ICC/IF image of ab76608 stained HepG2 cells. The cells were 100% Methanol fixed (5 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76608, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% Methanol fixed (5 min) MCF-7 cells at 5µg/ml.
  • IHC image of PDC6I staining in Human breast cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab76608, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-ALIX antibody (ab76608)

This product has been referenced in:
  • Corcoran C  et al. miR-34a is an intracellular and exosomal predictive biomarker for response to docetaxel with clinical relevance to prostate cancer progression. Prostate 74:1320-34 (2014). Read more (PubMed: 25053345) »

See 1 Publication for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"