For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    alix-antibody-epr23653-32-ab275377.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Cell Biology Apoptosis Extracellular Signals Death Ligands
Share by email
Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ALIX antibody [EPR23653-32] (ab275377)

  • Datasheet
  • SDS
  • Certificate of Compliance
Submit a review Submit a question References (1)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
  • Immunocytochemistry/ Immunofluorescence - Anti-ALIX antibody [EPR23653-32] (ab275377)
  • Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
  • Flow Cytometry (Intracellular) - Anti-ALIX antibody [EPR23653-32] (ab275377)
  • Immunoprecipitation - Anti-ALIX antibody [EPR23653-32] (ab275377)
  • Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
  • Flow Cytometry (Intracellular) - Anti-ALIX antibody [EPR23653-32] (ab275377)
  • Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
  • Immunocytochemistry/ Immunofluorescence - Anti-ALIX antibody [EPR23653-32] (ab275377)
  • Anti-ALIX antibody [EPR23653-32] (ab275377)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23653-32] to ALIX
  • Suitable for: WB, ICC/IF, Flow Cyt (Intra), IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Primary
Product image
Anti-TSG101 antibody [EPR7130(B)] (ab125011)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Knockout
Product image
Human PDCD6IP knockout HEK-293 cell line (ab260864)

View more associated products

Overview

  • Product name

    Anti-ALIX antibody [EPR23653-32]
    See all ALIX primary antibodies
  • Description

    Rabbit monoclonal [EPR23653-32] to ALIX
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt (Intra), IPmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: C6, RAW 264.7, PC-12, NIH/3T3, K-562, HEK-293, HeLa, HCT116, MCF7 and Jurkat whole cell lysates; Human brain tissue lysate; Mouse brain tissue lysate; Rat brain tissue lysate. ICC/IF: NIH/3T3 and HeLa cells. Flow Cyt (intra): NIH/3T3 and HeLa cells. IP: K562 whole cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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: 59.94% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR23653-32
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Anti-ALIX antibody [EPR23653-32] - BSA and Azide free (ab275387)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab275377 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
1/1000. Detects a band of approximately 80, 90, 100 kDa (predicted molecular weight: 96 kDa).
ICC/IF
1/5000.
Flow Cyt (Intra)
1/500.
IP
1/30.
Notes
WB
1/1000. Detects a band of approximately 80, 90, 100 kDa (predicted molecular weight: 96 kDa).
ICC/IF
1/5000.
Flow Cyt (Intra)
1/500.
IP
1/30.
Application notes
Is unsuitable for IHC-P.

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 [EPR23653-32] (ab275377)
    Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
    All lanes : Anti-ALIX antibody [EPR23653-32] (ab275377) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293 cell lysate
    Lane 2 : PDCD6IP knockout HEK-293 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Human Spleen tissue lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 96 kDa
    Observed band size: 96 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab275377 observed at 96 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab275377 was shown to react with PDCD6IP in wild-type HEK-293T cells in Western blot with loss of signal observed in PDCD6IP knockout cell line ab260864 (PDCD6IP knockout cell lysate ab261656). Wild-type HEK-293T and PDCD6IP knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5 % milk in TBS-T (0.1 % Tween®) before incubation with ab275377 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-ALIX antibody [EPR23653-32] (ab275377)
    Immunocytochemistry/ Immunofluorescence - Anti-ALIX antibody [EPR23653-32] (ab275377)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling ALIX with ab275377 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488, ab150077) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cells is observed. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594, ab195889) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488, ab150077) at 1/1000 dilution.

  • Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
    Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
    All lanes : Anti-ALIX antibody [EPR23653-32] (ab275377) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate
    Lane 3 : C6 (rat glial tumor glial cell) whole cell lysate
    Lane 4 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 5 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 6 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 96 kDa
    Observed band size: 80/90/100 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression pattern is consistent with what has been described in the literature (PMID: 24834918, 26935291, 28322231).

    Exposure time: Lane1-2: 10 seconds Lane3-6: 8 seconds.

     

  • Flow Cytometry (Intracellular) - Anti-ALIX antibody [EPR23653-32] (ab275377)
    Flow Cytometry (Intracellular) - Anti-ALIX antibody [EPR23653-32] (ab275377)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell line) cells labelling ALIX with ab275377 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). 

    A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

     

     

  • Immunoprecipitation - Anti-ALIX antibody [EPR23653-32] (ab275377)
    Immunoprecipitation - Anti-ALIX antibody [EPR23653-32] (ab275377)

    ALIX was immunoprecipitated from 0.35 mg K-562 (human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate with ab275377 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275377 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: K-562 whole cell lysate 10 ug

    Lane 2: ab275377 IP in K-562 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275377 in K-562 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds

  • Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
    Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
    Anti-ALIX antibody [EPR23653-32] (ab275377) at 1/1000 dilution + Human brain tissue lysate at 20 µg

    Secondary
    VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

    Predicted band size: 96 kDa
    Observed band size: 80/90/100 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression pattern is consistent with what has been described in the literature (PMID: 24834918, 26935291, 28322231).

    Exposure time: 10 seconds.

     

  • Flow Cytometry (Intracellular) - Anti-ALIX antibody [EPR23653-32] (ab275377)
    Flow Cytometry (Intracellular) - Anti-ALIX antibody [EPR23653-32] (ab275377)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) cells labelling ALIX with ab275377 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). 

    A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

     

     

  • Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
    Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)
    All lanes : Anti-ALIX antibody [EPR23653-32] (ab275377) at 1/1000 dilution

    Lane 1 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
    Lane 2 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate
    Lane 5 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
    Lane 6 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 96 kDa
    Observed band size: 90/100 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression pattern is consistent with what has been described in the literature (PMID: 24834918, 26935291, 28322231).

    Exposure time: 10 seconds.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-ALIX antibody [EPR23653-32] (ab275377)
    Immunocytochemistry/ Immunofluorescence - Anti-ALIX antibody [EPR23653-32] (ab275377)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embyro fibroblast cell line) cells labelling ALIX with ab275377 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488, ab150077 ) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cells. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594, ab195889) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Anti-ALIX antibody [EPR23653-32] (ab275377)
    Anti-ALIX antibody [EPR23653-32] (ab275377)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (1)

Publishing research using ab275377? Please let us know so that we can cite the reference in this datasheet.

ab275377 has been referenced in 1 publication.

  • Zhang R  et al. Exosomal SNHG16 secreted by CSCs promotes glioma development via TLR7. Stem Cell Res Ther 12:349 (2021). PubMed: 34134771

Customer reviews and Q&As

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

There are currently no Customer reviews or Questions for ab275377.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Get resources and offers direct to your inbox Sign up
A-Z by research area
  • Cancer
  • Cardiovascular
  • Cell biology
  • Developmental biology
  • Epigenetics & Nuclear signaling
  • Immunology
  • Metabolism
  • Microbiology
  • Neuroscience
  • Signal transduction
  • Stem cells
A-Z by product type
  • Primary antibodies
  • Secondary antibodies
  • Biochemicals
  • Isotype controls
  • Flow cytometry multi-color selector
  • Kits
  • Loading controls
  • Lysates
  • Peptides
  • Proteins
  • Slides
  • Tags and cell markers
  • Tools & Reagents
Help & support
  • Support
  • Make an Inquiry
  • Protocols & troubleshooting
  • Placing an order
  • RabMAb products
  • Biochemical product FAQs
  • Training
  • Browse by Target
Company
  • Corporate site
  • Investor relations
  • Company news
  • Careers
  • About us
  • Blog
Events
  • Tradeshows
  • Conferences
International websites
  • abcam.cn
  • abcam.co.jp

Join with us

  • LinkedIn
  • facebook
  • Twitter
  • YouTube
  • Terms of sale
  • Website terms of use
  • Cookie policy
  • Privacy policy
  • Legal
  • Modern slavery statement
© 1998-2023 Abcam plc. All rights reserved.