Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-ALIX antibody [EPR23653-32] - BSA and Azide free
See all ALIX primary antibodies
Description

Rabbit monoclonal [EPR23653-32] to ALIX - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC	Human
IP	Human
WB	Human

See all applications and species data

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: NIH/3T3 and HeLa cells. Flow cyt: NIH/3T3 and HeLa cells. IP: K562 whole cell lysate.
General notes
ab275387 is the carrier-free version of ab275377. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab275387 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
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.
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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C.
Storage buffer

Constituent: 100% PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23653-32
Isotype

IgG
Research areas

Immunocytochemistry - Anti-ALIX antibody [EPR23653-32] - BSA and Azide free (ab275387)

This data was developed using ab275377, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275387)

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?

This data was developed using ab275377, the same antibody clone in a different buffer formulation.

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

This data was developed using ab275377, the same antibody clone in a different buffer formulation.

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) (Black) isotype control 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] - BSA and Azide free (ab275387)

This data was developed using ab275377, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275387)

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?

This data was developed using ab275377, the same antibody clone in a different buffer formulation.

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

This data was developed using ab275377, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (ab275387)

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?

This data was developed using ab275377, the same antibody clone in a different buffer formulation.

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

This data was developed using ab275377, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic 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] - BSA and Azide free (ab275387)