Product nameAnti-Cyclophilin A antibody
See all Cyclophilin A primary antibodies
DescriptionMouse monoclonal to Cyclophilin A
Tested applicationsSuitable for: WB, ICC/IF, ELISA, IP, Flow Cytmore details
Species reactivityReacts with: Human
Recombinant full length protein, corresponding to amino acids 1-166 of Human Cyclophilin A
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferPreservative: None
PBS, pH 7.2
Concentration information loading...
PurityProtein G purified
Light chain typekappa
Our Abpromise guarantee covers the use of ab58144 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 - 5 µg/ml. Predicted molecular weight: 18 kDa.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|ELISA||Use at an assay dependent concentration. It can be applied in indirect ELISA (at a dilution of 1/1500 for 24 hrs).|
|IP||Use at an assay dependent concentration.|
|Flow Cyt||Use 0.5µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
FunctionPPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides.
Sequence similaritiesBelongs to the cyclophilin-type PPIase family. PPIase A subfamily.
Contains 1 PPIase cyclophilin-type domain.
- Information by UniProt
- Cyclophilin A antibody
- Cyclophilin antibody
- CyclophilinA antibody
Cyclophilin A antibody (ab58144) at 1ug/lane + Jurkat cell lysate at 25ug/lane.
ICC/IF image of ab58144 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab58144, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Cyclophilin A was immunoprecipitated using 0.5mg Hela whole cell extract, 10µg of Mouse monoclonal to Cyclophilin A and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab58144.
Secondary: Protein G-HRP at 1/500 dilution.
Band: 18kDa: Cyclophilin A.
Overlay histogram showing HeLa cells stained with ab58144 (red line). The cells were fixed with 4% paraformaldehyde 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 followed by the antibody (ab58144, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This image shows an ELISA analysis of ab58144.
PBMC huh-7 cells were incubated with medium + 10% FCS. At the end of the incubation the supernatant was collected and cells washed with cold PBS. Cells were scrapped and lysate generated. The cell lysate and extracellular medium were assyed for Cyclphilin A content in a sandwich ELISA. The primary antibody was diluted 1/1500 and incubated with the sample for 24 hours at 4°C. Ab6729 was diluted 1/2000.
Key for lower diagram:
1) Rabbit anti-Cyclophilin A (ab3563)
2) Cells samples or Std (ab56523)
3) Mouse anti-Cyclophilin A (ab58144)
4) Rabbit anti-mouse Alkaline Phosphatase (ab6729)
5) Substrate of Alkaline Phosphatase
Standards = Recombinant protein Cyclophilin A (ab56523)
This product has been referenced in:
- Li T et al. Ubiquitin-specific protease 4 promotes hepatocellular carcinoma progression via cyclophilin A stabilization and deubiquitination. Cell Death Dis 9:148 (2018). Read more (PubMed: 29396555) »
- Hirschmugl B et al. Maternal obesity modulates intracellular lipid turnover in the human term placenta. Int J Obes (Lond) 41:317-323 (2017). WB ; Human . Read more (PubMed: 27780978) »