Key features and details
- Rabbit polyclonal to CHMP2B
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Product nameAnti-CHMP2B antibody
See all CHMP2B primary antibodies
DescriptionRabbit polyclonal to CHMP2B
Tested applicationsSuitable for: WB, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Chicken
Synthetic peptide corresponding to Human CHMP2B aa 150 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available as
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In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
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.
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 24 kDa.|
|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 has been referenced in 31 publications.
- Streetley J et al. Stimulated release of intraluminal vesicles from Weibel-Palade bodies. Blood 133:2707-2717 (2019). PubMed: 30760452
- Tabata K et al. Immuno-localization of ESCRT Proteins in Virus-Infected Cells by Fluorescence and Electron Microscopy. Methods Mol Biol 1998:73-92 (2019). PubMed: 31250295
- Kumar H et al. The exocyst complex and Rab5 are required for abscission by localizing ESCRT III subunits to the cytokinetic bridge. J Cell Sci 132:N/A (2019). PubMed: 31221728
- Adar-Levor S et al. Studying the Spatial Organization of ESCRTs in Cytokinetic Abscission Using the High-Resolution Imaging Techniques SIM and Cryo-SXT. Methods Mol Biol 1998:129-148 (2019). PubMed: 31250299
- Wiersma VI et al. Granulovacuolar degeneration bodies are neuron-selective lysosomal structures induced by intracellular tau pathology. Acta Neuropathol 138:943-970 (2019). PubMed: 31456031