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Rabbit Recombinant Monoclonal ALIX antibody. Suitable for ICC/IF, IP, WB, Flow Cyt (Intra) and reacts with Mouse, Human, Rat samples. Cited in 29 publications.


Images

Western blot - Anti-ALIX antibody [EPR23653-32] (AB275377), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-ALIX antibody [EPR23653-32] (AB275377), expandable thumbnail
  • Immunoprecipitation - Anti-ALIX antibody [EPR23653-32] (AB275377), expandable thumbnail
  • Western blot - Anti-ALIX antibody [EPR23653-32] (AB275377), expandable thumbnail
  • Western blot - Anti-ALIX antibody [EPR23653-32] (AB275377), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFIPWBIHC-PFlow Cyt (Intra)
Human
Tested
Tested
Tested
Not recommended
Tested
Mouse
Tested
Expected
Tested
Not recommended
Tested
Rat
Not recommended
Expected
Tested
Not recommended
Not recommended

Tested
Tested

Species
Mouse
Dilution info
1/5000
Notes

-

Species
Human
Dilution info
1/5000
Notes

-

Not recommended
Not recommended

Species
Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Rat
Dilution info
1/1000
Notes

-

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/500
Notes

-

Species
Human
Dilution info
1/500
Notes

-

Not recommended
Not recommended

Species
Rat
Dilution info
-
Notes

-

Target data

Function

Multifunctional protein involved in endocytosis, multivesicular body biogenesis, membrane repair, cytokinesis, apoptosis and maintenance of tight junction integrity. 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 requires the sequential function of ESCRT-O, -I,-II and -III complexes (PubMed:14739459). The ESCRT machinery also functions in topologically equivalent membrane fission events, such as the terminal stages of cytokinesis (PubMed:17556548, PubMed:17853893). Adapter for a subset of ESCRT-III proteins, such as CHMP4, to function at distinct membranes. Required for completion of cytokinesis (PubMed:17556548, PubMed:17853893, PubMed:18641129). May play a role in the regulation of both apoptosis and cell proliferation. Regulates exosome biogenesis in concert with SDC1/4 and SDCBP (PubMed:22660413). By interacting with F-actin, PARD3 and TJP1 secures the proper assembly and positioning of actomyosin-tight junction complex at the apical sides of adjacent epithelial cells that defines a spatial membrane domain essential for the maintenance of epithelial cell polarity and barrier (By similarity). (Microbial infection) Involved in HIV-1 virus budding. Can replace TSG101 it its role of supporting HIV-1 release; this function requires the interaction with CHMP4B. The ESCRT machinery also functions in topologically equivalent membrane fission events, such as enveloped virus budding (HIV-1 and other lentiviruses).

Alternative names

AIP1, ALIX, KIAA1375, PDCD6IP, Programmed cell death 6-interacting protein, PDCD6-interacting protein, ALG-2-interacting protein 1, ALG-2-interacting protein X, Hp95

Recommended products

Rabbit Recombinant Monoclonal ALIX antibody. Suitable for ICC/IF, IP, WB, Flow Cyt (Intra) and reacts with Mouse, Human, Rat samples. Cited in 29 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR23653-32
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

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 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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

ALIX also known as PDCD6IP or ALIX G is an adapter protein involved in various cellular processes. It holds a molecular weight of approximately 95-100 kDa and is widely expressed in different tissue types including HEK 293 cells. ALIX plays a significant role as a component of the endosomal sorting complexes required for transport (ESCRT) machinery contributing to membrane dynamics and intracellular trafficking.

Biological function summary

ALIX acts as an important mediator in the regulation of cellular membrane remodeling. It associates with several protein complexes influencing the budding and fission of vesicles and endosomes. The protein also interacts with ubiquitin and other factors guiding protein sorting and maintaining cellular homeostasis. This adapter characteristic makes ALIX essential for recycling receptors and maintaining cell surface receptor expression levels.

Pathways

ALIX is involved in the ESCRT pathway and the apoptotic pathways. It works closely with proteins like TSG101 and CHMP4 to facilitate vesicle formation and dispatch impacting cellular processes like signaling and waste disposal. Furthermore ALIX regulates apoptosis by interacting with proteins such as ALG-2 and to some extent with apoptosis-related membranes to mediate programmed cell death.

Associated diseases and disorders

ALIX has established links to cancer progression and neurodegenerative disorders. Its participation in membrane remodeling and protein sorting correlates with tumor development with evidence showing that altered expression affects oncogenic pathways. Additionally dysregulation of ALIX-associated pathways has implications in neurodegenerative diseases where ALIX and proteins such as those in the ESCRT complex could play a role in the degradation of misfolded proteins and endocytic dysfunctions contributing to disease pathology.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377), expandable thumbnail

    Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)

    Lanes 1 - 4: Merged signal (red and green). Green - ab275377 observed at 96 kDa. Red - loading control Anti-GAPDH antibody [6C5] - 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 Human PDCD6IP knockout HEK-293 cell line ab260864 (PDCD6IP knockout cell lysate Human PDCD6IP knockout HEK-293 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

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

    Lane 1: Wild-type HEK-293 cell lysate at 20 µg

    Lane 2: PDCD6IP knockout HEK-293 cell lysate at 20 µg

    Lane 3: HeLa cell lysate at 20 µg

    Lane 4: Human Spleen tissue lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 96 kDa

    Observed band size: 96 kDa

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

    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, 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, Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.

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

    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)(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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab275377 in K-562 whole cell lysate

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

    Exposure time: 3 seconds

    All lanes: Immunoprecipitation - Anti-ALIX antibody [EPR23653-32] (ab275377)

    Predicted band size: 96 kDa

    Observed band size: 90-100 kDa

  • Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377), expandable thumbnail

    Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)

    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.

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

    Lane 1: Mouse brain tissue lysate at 20 µg

    Lane 2: Rat brain tissue lysate at 20 µg

    Lane 3: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

    Lane 4: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

    Lane 5: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

    Lane 6: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

    Secondary

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

    Predicted band size: 96 kDa

    Observed band size: 100 kDa, 80 kDa, 90 kDa

  • Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377), expandable thumbnail

    Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)

    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.

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

    All lanes: Human brain tissue lysate at 20 µg

    Secondary

    All lanes: VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

    Predicted band size: 96 kDa

    Observed band size: 100 kDa, 80 kDa, 90 kDa

  • Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377), expandable thumbnail

    Western blot - Anti-ALIX antibody [EPR23653-32] (ab275377)

    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.

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

    Lane 1: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

    Lane 2: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Lane 3: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 5: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 6: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

    Secondary

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

    Predicted band size: 96 kDa

    Observed band size: 90-100 kDa

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

    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, 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, Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

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

    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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

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

    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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

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