The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Predicted molecular weight: 25 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
Use at an assay dependent concentration. PubMed: 21926173
Probable core component of the endosomal sorting required for transport complex III (ESCRT-III) which is involved in multivesicular bodies (MVBs) formation and sorting of endosomal cargo proteins into MVBs. MVBs contain intraluminal vesicles (ILVs) that are generated by invagination and scission from the limiting membrane of the endosome and mostly are delivered to lysosomes enabling degradation of membrane proteins, such as stimulated growth factor receptors, lysosomal enzymes and lipids. The MVB pathway appears to require the sequential function of ESCRT-O, -I,-II and -III complexes. ESCRT-III proteins mostly dissociate from the invaginating membrane before the ILV is released. The ESCRT machinery also functions in topologically equivalent membrane fission events, such as the terminal stages of cytokinesis and the budding of enveloped viruses (HIV-1 and other lentiviruses). ESCRT-III proteins are believed to mediate the necessary vesicle extrusion and/or membrane fission activities, possibly in conjunction with the AAA ATPase VPS4. When overexpressed, membrane-assembled circular arrays of CHMP4B filaments can promote or stabilize negative curvature and outward budding. Via its interaction with PDCD6IP involved in HIV-1 p6-and p9-dependent virus release.
Widely expressed. Expressed at higher level in heart and skeletal muscle. Also expressed in brain, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood lymphocytes.
Involvement in disease
Defects in CHMP4B are the cause of cataract posterior polar type 3 (CTPP3) [MIM:605387]. A subcapsular opacity, usually disk-shaped, located at the back of the lens. It can have a marked effect on visual acuity.
Belongs to the SNF7 family.
The acidic C-terminus and the basic N-termminus are thought to render the protein in a closed, soluble and inactive conformation through an autoinhibitory intramolecular interaction. The open and active conformation, which enables membrane binding and oligomerization, is achieved by interaction with other cellular binding partners, probably including other ESCRT components.
Vacuolar protein sorting-associated protein 32-2 antibody
Immunocytochemistry/ Immunofluorescence - Anti-CHMP4B antibody (ab105767)Image from Bodon G et al, J Biol Chem. 2011 Nov 18;286(46):40276-86. Epub 2011 Sep 16, Fig 3. DOI 10.1074/jbc.M111.283671
ab105767 staining CHMP4B (red) in HeLa cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with 4% paraformaldehyden and after three washes in PBS, cells were permeabilized with PBS containing 0.5% Triton X-100 in blocking solution (PBS containing 5% goat pre-immune serum) for 30 minutes. ab105767 was diluted in blocking solution and then incubated for 1 hour. After washing in PBS, cells were incubated for 1 hour with the secondary antibody.
Western blot - Anti-CHMP4B antibody (ab105767)
Anti-CHMP4B antibody (ab105767) at 1 µg/ml + lysate prepared from THP-1 cells