Product nameAnti-Separase antibody [XJ11-1B12]
See all Separase primary antibodies
DescriptionMouse monoclonal [XJ11-1B12] to Separase
Tested applicationsSuitable for: Flow Cyt, WB, ICC/IFmore details
Species reactivityReacts with: Human
Maltose binding protein fusion protein, corresponding to C terminal amino acids 1866-1996 of Human Separase.
- This antibody gave a positive signal in MDA-MB-231 Whole Cell Lysate.
This antibody clone is manufactured by Abcam.
Separase antibodies can be used as a specific marker for centrosomes of mitotic cells. The staining of separase in centrosomes can be detected from prophase of mitosis up until anaphase.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Concentration information loading...
Primary antibody notesSeparase antibodies can be used as a specific marker for centrosomes of mitotic cells. The staining of separase in centrosomes can be detected from prophase of mitosis up until anaphase.
Light chain typekappa
Our Abpromise guarantee covers the use of ab16170 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 10 µg/ml. Detects a band of approximately 170 kDa (predicted molecular weight: 233 kDa).|
|ICC/IF||Use at an assay dependent concentration. Used at a dilution of 1/100 for 1 hr on HeLa cells (see Abreview).|
FunctionCaspase-like protease, which plays a central role in the chromosome segregation by cleaving the SCC1/RAD21 subunit of the cohesin complex at the onset of anaphase. During most of the cell cycle, it is inactivated by different mechanisms.
Sequence similaritiesBelongs to the peptidase C50 family.
modificationsAutocleaves. This function, which is not essential for its protease activity, is unknown.
Phosphorylated by CDK1. There are 8 Ser/Thr phosphorylation sites. Among them, Ser-1126 phosphorylation is the major site, which conducts to the enzyme inactivation.
Cellular localizationCytoplasm. Nucleus.
- Information by UniProt
- Caspase like protein ESPL1 antibody
- Caspase-like protein ESPL1 antibody
- ESP 1 antibody
Anti-Separase antibody [XJ11-1B12] (ab16170) at 10 µg/ml + MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 20 µg
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 233 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa, 70 kDa (possible cleavage fragment). We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes
ab16170 stained MCF7 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16170 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Overlay histogram showing HeLa cells stained with ab16170 (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 followed by the antibody (ab16170, 1µg/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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This product has been referenced in:
- Karki M et al. Precocious centriole disengagement and centrosome fragmentation induced by mitotic delay. Nat Commun 8:15803 (2017). WB ; Human . Read more (PubMed: 28607478) »
- Seo MY et al. Integrity of the Pericentriolar Material Is Essential for Maintaining Centriole Association during M Phase. PLoS One 10:e0138905 (2015). WB . Read more (PubMed: 26407333) »