Anti-ALIX antibody [3A9] (ab117600)

Mouse monoclonal ALIX antibody [3A9]. Validated in WB, IP, ELISA, IHC, ICC/IF and tested in Mouse, Rat, Human, Xenopus laevis, Xenopus tropicalis. Cited in 16 publication(s).

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: WB, ELISA, IHC-P, ICC/IF, IPmore details
  • Species reactivity

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

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

  • Positive control

Properties

Applications

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 Use at an assay dependent concentration. Predicted molecular weight: 96 kDa.
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Do not perform antigen retrieval.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

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

  • 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

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

    Secondary
    Rabbit Anti Mouse IgG

    Predicted band size: 96 kDa

References

This product has been referenced in:

See all 19 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Aug 20 2014

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

Abcam user community

Verified customer

Submitted Aug 20 2014

Question
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.

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