Mouse, Rat, Rabbit, Guinea pig, Hamster, Cow, Dog, Human, Pig, Saccharomyces cerevisiae, Caenorhabditis elegans, Drosophila melanogaster, Monkey Predicted to work with:
Recombinant Mouse TCP1 alpha protein fragment (carboxy terminal region).
EpitopeThis affinity purified antibody recognizes an epitope in the C terminus
of mouse TCP1 alpha (465-AKLRA).
HeLa, PC12 and 3T3 and RK-13 cell lysates (all heat shocked).
This antibody gave a positive result in IHC in the following FFPE tissue: Human testis seminoma.
ab90357 reacts weakly with Saccharomyces cerevisiae, consistent with the epitope sequence being AKLRS (instead of AKLRA). In C. elegans, ab90357 reacts with TCP1 alpha and another CCT subunit. In plants, ab90357 recognizes TCP1 of Pisum sativum, and the sequence of Arabidopsis thalania TCP1 over the region of the epitope AKLRA. It has also been shown that ab90357 reacts with a subunit of a specialized chaperonin which folds phytochrome.
Storage instructionsShipped at 4°C. Store at -20ºC.
Primary antibody notesab90357 reacts weakly with Saccharomyces cerevisiae, consistent with the epitope sequence being AKLRS (instead of AKLRA). In C. elegans, ab90357 reacts with TCP1 alpha and another CCT subunit. In plants, ab90357 recognizes TCP1 of Pisum sativum, and the sequence of Arabidopsis thalania TCP1 over the region of the epitope AKLRA. It has also been shown that ab90357 reacts with a subunit of a specialized chaperonin which folds phytochrome.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Predicted molecular weight: 60 kDa.
Use at an assay dependent concentration.
Use 1µg for 106 cells. ab18450-Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
FunctionMolecular chaperone; assists the folding of proteins upon ATP hydrolysis. As part of the BBS/CCT complex may play a role in the assembly of BBSome, a complex involved in ciliogenesis regulating transports vesicles to the cilia. Known to play a role, in vitro, in the folding of actin and tubulin.
Sequence similaritiesBelongs to the TCP-1 chaperonin family.
IHC image of TCP1 alpha staining in Human testis seminoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab90357, 5µg/ml, for 15 mins at room temperature. A Goat anti-Rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Overlay histogram showing HeLa cells stained with ab90357 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab90357, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] ab18540, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
References for Anti-TCP1 alpha antibody [91a] (ab90357)
This product has been referenced in:
Noormohammadi A et al. Somatic increase of CCT8 mimics proteostasis of human pluripotent stem cells and extends C. elegans lifespan. Nat Commun7:13649 (2016).
Human, Caenorhabditis elegans
Read more (PubMed: 27892468) »
Monzo K et al. Proteomic analysis reveals CCT is a target of Fragile X mental retardation protein regulation in Drosophila. Dev Biol340:408-18 (2010).
Read more (PubMed: 20122915) »