Anti-ALIX antibody (ab88388)
Key features and details
- Rabbit polyclonal to ALIX
- Suitable for: WB, ICC
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
-
Product name
Anti-ALIX antibody
See all ALIX primary antibodies -
Description
Rabbit polyclonal to ALIX -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC HumanWB Human
-
Immunogen
Synthetic peptide corresponding to Human ALIX aa 300-400 conjugated to keyhole limpet haemocyanin.
(Peptide available asab89369) -
Positive control
- This antibody gave a positive signal in the following whole cell lysates: HeLa; HepG2; HEK-293, SHSY5Y; HUVEC. ICC: Hek293 cell line ICC KO: HEK293 (HEK293-ALIX KO used as a negative cell line)
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab88388 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC HumanWB HumanAll applications MouseRatOrangutanApplication Abreviews Notes WB (2) Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 96 kDa).ICC Use a concentration of 5 µg/ml.Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 96 kDa).ICC
Use a concentration of 5 µg/ml.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. - 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
see all -
Alternative names
- AIP1 antibody
- ALG 2 interacting protein 1 antibody
- ALG-2-interacting protein 1 antibody
see all
Images
-
Immunocytochemistry - Anti-ALIX antibody (ab88388)Lab
ab88388 staining ALIX in Hek293 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab88388 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
-
Immunocytochemistry - Anti-ALIX antibody (ab88388)ab88388 staining ALIX in wild-type HEK293 cells (top panel) and ALIX knockout HEK293 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab88388 at 5µg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).
-
Western blot - Anti-ALIX antibody (ab88388)All lanes : Anti-ALIX antibody (ab88388) at 1 µg/ml
Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : ALIX knockout HEK-293 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 96 kDa
Observed band size: 96 kDaLanes 1 - 3: Merged signal (red and green). Green - ab88388 observed at 96 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab88388 was shown to recognize in wild-type HEK-293 cells as signal was lost at the expected MW in ALIX knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and ALIX knockout samples were subjected to SDS-PAGE. ab88388 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
-
Western blot - Anti-ALIX antibody (ab88388)All lanes : Anti-ALIX antibody (ab88388) 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
All lanes : 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: 44 kDa, 82 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
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 than predicted.
Protocols
Datasheets and documents
References (5)
ab88388 has been referenced in 5 publications.
- Uretmen Kagiali ZC et al. CLIC4 and CLIC1 bridge plasma membrane and cortical actin network for a successful cytokinesis. Life Sci Alliance 3:N/A (2020). PubMed: 31879279
- Thepparit C et al. Dengue virus requires apoptosis linked gene-2-interacting protein X (ALIX) for viral propagation. Virus Res 261:65-71 (2019). PubMed: 30599162
- Skalnikova HK et al. Isolation and Characterization of Small Extracellular Vesicles from Porcine Blood Plasma, Cerebrospinal Fluid, and Seminal Plasma. Proteomes 7:N/A (2019). PubMed: 31027284
- Longatti A et al. High affinity single-chain variable fragments are specific and versatile targeting motifs for extracellular vesicles. Nanoscale 10:14230-14244 (2018). PubMed: 30010165
- Zheng T et al. Plasma Exosomes Spread and Cluster Around ß-Amyloid Plaques in an Animal Model of Alzheimer's Disease. Front Aging Neurosci 9:12 (2017). WB . PubMed: 28203202
Images
-
Immunocytochemistry - Anti-ALIX antibody (ab88388) Lab
ab88388 staining ALIX in Hek293 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab88388 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
-
Immunocytochemistry - Anti-ALIX antibody (ab88388)
ab88388 staining ALIX in wild-type HEK293 cells (top panel) and ALIX knockout HEK293 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab88388 at 5µg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).
-
Western blot - Anti-ALIX antibody (ab88388)All lanes : Anti-ALIX antibody (ab88388) at 1 µg/ml
Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : ALIX knockout HEK-293 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 96 kDa
Observed band size: 96 kDaLanes 1 - 3: Merged signal (red and green). Green - ab88388 observed at 96 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab88388 was shown to recognize in wild-type HEK-293 cells as signal was lost at the expected MW in ALIX knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and ALIX knockout samples were subjected to SDS-PAGE. ab88388 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
-
Western blot - Anti-ALIX antibody (ab88388)All lanes : Anti-ALIX antibody (ab88388) 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
All lanes : 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: 44 kDa, 82 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
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 than predicted.
