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    alix-antibody-3a9-ab117600.pdf

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Cell Biology Apoptosis Extracellular Signals Death Ligands
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Anti-ALIX antibody [3A9] (ab117600)

  • Datasheet
  • SDS
Reviews (3)Q&A (1)References (68)

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Western blot - Anti-ALIX antibody [3A9] (ab117600)

    Key features and details

    • Mouse monoclonal [3A9] to ALIX
    • Suitable for: WB
    • Reacts with: Human
    • Isotype: IgG1

    You may also be interested in

    Protein
    Product image
    Recombinant Human ALIX protein (ab132534)
    Secondary
    Product image
    Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    Primary
    Product image
    Anti-CD63 antibody [MX-49.129.5] - BSA and Azide free (ab213092)

    View more associated products

    Overview

    • Product name

      Anti-ALIX antibody [3A9]
      See all ALIX primary antibodies
    • Description

      Mouse monoclonal [3A9] to ALIX
    • Host species

      Mouse
    • Tested applications

      Suitable for: WBmore details
    • Species reactivity

      Reacts with: Human
      Predicted to work with: Mouse, Rat
    • Immunogen

      Fusion protein (GST-tag) corresponding to Human ALIX aa 600-750 (N terminal).
      Database link: Q8WUM4

    • Positive control

      • WB: HeLa whole cell lysate (ab150035)
    • 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. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
    • Storage buffer

      Preservative: 0.09% Sodium azide
      Constituent: 99% PBS
    • Concentration information loading...
    • Purity

      Protein G purified
    • Clonality

      Monoclonal
    • Clone number

      3A9
    • Isotype

      IgG1
    • Research areas

      • Cell Biology
      • Apoptosis
      • Extracellular Signals
      • Death Ligands
      • Microbiology
      • Interspecies Interaction
      • Host Virus Interaction
      • Signal Transduction
      • Protein Trafficking
      • Vesicle Transport
      • Regulation

    Associated products

    • Compatible Secondaries

      • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
      • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Isotype control

      • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Positive Controls

      • HeLa whole cell lysate (ab29545)
    • Recombinant Protein

      • Recombinant Human ALIX protein (ab132534)

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab117600 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 (2)
    Use at an assay dependent concentration. Predicted molecular weight: 96 kDa.
    Notes
    WB
    Use at an assay dependent concentration. Predicted molecular weight: 96 kDa.

    Target

    • Function

      Class 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 similarities

      Contains 1 BRO1 domain.
    • Cellular localization

      Cytoplasm > 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.
    • Target information above from: UniProt accession Q8WUM4 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 10015 Human
      • Entrez Gene: 18571 Mouse
      • Entrez Gene: 501083 Rat
      • Omim: 608074 Human
      • SwissProt: Q8WUM4 Human
      • SwissProt: Q9WU78 Mouse
      • SwissProt: Q9QZA2 Rat
      • Unigene: 475896 Human
      • Unigene: 29816 Mouse
      • Unigene: 101381 Rat
      • Unigene: 1588 Rat
      • Unigene: 226240 Rat
      see all
    • 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

    Images

    • Western blot - Anti-ALIX antibody [3A9] (ab117600)
      Western blot - Anti-ALIX antibody [3A9] (ab117600)
      Anti-ALIX antibody [3A9] (ab117600) + HeLa whole cell lysate

      Secondary
      Rabbit Anti Mouse IgG

      Predicted band size: 96 kDa

    Protocols

    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (68)

    Publishing research using ab117600? Please let us know so that we can cite the reference in this datasheet.

    ab117600 has been referenced in 68 publications.

    • Lázaro-Ibáñez E  et al. Selection of Fluorescent, Bioluminescent, and Radioactive Tracers to Accurately Reflect Extracellular Vesicle Biodistribution in Vivo. ACS Nano 15:3212-3227 (2021). PubMed: 33470092
    • Zhang Z  et al. Human umbilical cord mesenchymal stem cell-derived exosomal miR-146a-5p reduces microglial-mediated neuroinflammation via suppression of the IRAK1/TRAF6 signaling pathway after ischemic stroke. Aging (Albany NY) 13:3060-3079 (2021). PubMed: 33479185
    • Chhoy P  et al. Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models. STAR Protoc 2:100303 (2021). PubMed: 33554138
    • Valdes R  et al. Cellular Trafficking of Glutathione Transferase M2-2 Between U373MG and SHSY-S7 Cells is Mediated by Exosomes. Neurotox Res 39:182-190 (2021). PubMed: 33555546
    • Qin Z  et al. miR-224-5p Contained in Urinary Extracellular Vesicles Regulates PD-L1 Expression by Inhibiting Cyclin D1 in Renal Cell Carcinoma Cells. Cancers (Basel) 13:N/A (2021). PubMed: 33557163
    View all Publications for this product

    Customer reviews and Q&As

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

    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-ALIX antibody [3A9]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human Cervical cancer cell lines)
    Gel Running Conditions
    Reduced Denaturing (8% SDS page gel)
    Loading amount
    30 µg
    Treatment
    10uM CCCP for 24h
    Specification
    Human Cervical cancer cell lines
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 2-8°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 Nov 18 2019

    Western blot abreview for Anti-ALIX antibody [3A9]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (Epethelial cells)
    Specification
    Epethelial cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°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 Aug 20 2014

    Immunoprecipitation abreview for Anti-ALIX antibody [3A9]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Immuno-precipitation step
    Protein A/G
    Sample
    Human Cell lysate - whole cell (Epethelial cells)
    Specification
    Epethelial cells
    Total protein in input
    250 µg
    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 Aug 20 2014

    Question

    send WB protocol (Ab dilution etc.)

    Read More

    Abcam community

    Verified customer

    Asked on May 15 2012

    Answer

    Thank you for contacting us.

    Here is the WB protocol you had requested.

    Western Blotting

    Reagents
    1. Blocking Buffer
    5% non-fat dried milk dissolved in PBS.
    Note: For the detection of phosphoproteins, NEVER block membranes with milk; use 5% BSA instead. Milk contains casein (itself a phosphoprotein) which will bind phospho-specific antibodies resulting in high background.
    2. Washing Buffer
    Blocking buffer + 0.1% Tween-20
    3. Ponceau S
    Ponceau S, 0.1 g
    Acetic Acid, 5 ml
    Distilled Water, 95 ml
    Note: Ponceau S is light sensitive.
    4. PBS
    Sodium Chloride, 8.0g Potassium Chloride, 0.2g
    Disodium Potassium Phosphate, 1.15g
    Potassium Dihydrogen Phosphate, 0.2g
    Distilled Water, 1 liter
    5. PBST
    Sodium Chloride, 8.0g
    Potassium Chloride, 0.2g
    Disodium Potassium Phosphate, 1.15g
    Potassium Dihydrogen Phosphate, 0.2g
    Tween-20, 1.0ml
    Distilled Water, 1 liter
    Method
    Following SDS-PAGE, transfer proteins onto blotting membrane according to the manufacturer’s instructions.
    Check protein transfer by staining the blot with Ponceau S for 1 minute, then completely destain the blot by washing with distilled water.
    Place blot into blocking solution for 2 hours at room temperature, or overnight at 4°C.
    Rinse the blot briefly with wash buffer and then add primary antibody diluted in the wash buffer (a concentration of 1-10 µg/ml is generally acceptable and recommended). Incubate for 2 hours at room temperature, or overnight at 4°C.
    Wash the blot extensively in wash buffer (3 x 10 minutes) with gentle agitation.
    Add appropriate enzyme-conjugated secondary antibody (e.g. ab97023 (https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-HL-HRP-ab97023.html) or ab97046 (https://www.abcam.com/Rabbit-polyclonal-Secondary-Antibody-to-Mouse-IgG-H-L-HRP-ab97046.html)) diluted in wash buffer and incubate for 1 hour at room temperature with gentle agitation.
    Wash the membrane with gentle agitation as follows: 4 x 5 minutes in wash buffer; 3 x 5 minutes in PBST and 2 x 5 minutes in PBS.
    Add appropriate enzyme substrate solution and incubate as recommended by the manufacturer to visualize protein bands.

    Appropriate controls should always be carried out. It may be useful to include a sample in which no primary antibody is used at all, in order to determine any non-specific binding of the secondary reagent to the target tissue.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on May 15 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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