Product nameAnti-CHMP2B antibody
See all CHMP2B primary antibodies
DescriptionRabbit polyclonal to CHMP2B
Tested applicationsSuitable for: IHC-FoFr, WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Chicken
Synthetic peptide corresponding to Human CHMP2B aa 150 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available as
- WB: Tagged recombinant CHMP2B protein, Wild-type A549 whole cell lysate, A431 whole cell lysate, HeLa whole cell lysate
Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab33174 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-FoFr||Use at an assay dependent concentration. See Abreview.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 24 kDa.|
|IHC-P||Use at an assay dependent concentration. PubMed: 20420883|
|ICC/IF||Use a concentration of 5 µg/ml.|
FunctionProbable 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.
Tissue specificityWidely expressed. Expressed in brain, heart, skeletal muscle, spleen, kidney, liver, small intestine, pancreas, lung, placenta and leukocytes. In brain, it is expressed in cerebellum, cerebral cortex, medulla, spinal chord, occipital lobe, frontal lobe, temporal lobe and putamen.
Involvement in diseaseDefects in CHMP2B are the cause of frontotemporal dementia, chromosome 3-linked (FTD3) [MIM:600795]. FTD3 is characterized by an onset of dementia in the late 50's initially characterized by behavioral and personality changes including apathy, restlessness, disinhibition and hyperorality, progressing to stereotyped behaviors, non-fluent aphasia, mutism and dystonia, with a marked lack of insight. The brains of individuals with FTD3 have no distinctive neuropathological features. They show global cortical and central atrophy, but no beta-amyloid deposits.
Sequence similaritiesBelongs to the SNF7 family.
DomainThe 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.
Cellular localizationCytoplasm > cytosol. Late endosome membrane.
- Information by UniProt
- ALS17 antibody
- Charged multivesicular body protein 2b antibody
- CHM2B_HUMAN antibody
All lanes : Anti-CHMP2B antibody (ab33174) at 1 µg/ml
Lane 1 : Wild-type A549 whole cell lysate
Lane 2 : CHMP2B knockout A549 whole cell lysate
Lane 3 : A431 whole cell lysate
Lane 4 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 24 kDa
Lanes 1 - 4: Merged signal (red and green). Green - ab33174 observed at 30 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab33174 was shown to recognize CHMP2B in wild-type A549 cells as signal was lost at the expected MW in CHMP2B knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CHMP2B knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab33174 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/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.
Anti-CHMP2B antibody (ab33174) at 1 µg/ml + Tagged recombinant CHMP2B protein at 0.1 µg
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 24 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?
Ab33174 was tested on the full length recombinant tagged CHMP2B protein which is predicted to run at 50 kDa.
ICC/IF image of ab33174 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab33174, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
ab33174 staining CHMP2B in mouse brain tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue from 4% PFA perfused animals underwent overnight fixation in 4% paraformaldehyde, cryoprotected in 30% sucrose and cut using cryostat.The primary antibody was diluted, 1/300 (PBS + 0.3% Triton X100) and incubated with sample for 18 hours at 20°C. An abcam antibody ab60314, Chromeo488 conjugated goat polyclonal to rabbit IgG, diluted 1/1000 was used as secondary. The antibody produced a diffuse and widespread staining in many brain areas. The image shows the staining obtained in the mouse hippocampus.
ab33174 staining CHMP2B in human hippocampus tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then blocked with 4% serum for 1 hour at 27°C followed by incubation with the primary antibody at a 1/600 dilution for 18 hours at 4°C. A Cy3®-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/500 dilution.
This product has been referenced in:
- Streetley J et al. Stimulated release of intraluminal vesicles from Weibel-Palade bodies. Blood 133:2707-2717 (2019). Read more (PubMed: 30760452) »
- Zhang L et al. Common Deregulation of Seven Biological Processes by MicroRNAs in Gastrointestinal Cancers. Sci Rep 8:3287 (2018). WB ; Human . Read more (PubMed: 29459716) »