Anti-Separase antibody [XJ11-1B12] (ab16170)
![Anti-Separase antibody [XJ11-1B12] (ab16170)](https://www.abcam.com/ps/products/16/ab16170/Images/ab16170-24880-anti-separase-antibody-xj11-1b12-western-blot.jpg "Anti-Separase antibody [XJ11-1B12] (ab16170)")
Key features and details
- Mouse monoclonal [XJ11-1B12] to Separase
- Suitable for: Flow Cyt, WB, ICC/IF
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-Separase antibody [XJ11-1B12]
See all Separase primary antibodies -
Description
Mouse monoclonal [XJ11-1B12] to Separase -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- This antibody gave a positive signal in MDA-MB-231 Whole Cell Lysate.
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General notes
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.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Protein G purified -
Primary antibody notes
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. -
Clonality
Monoclonal -
Clone number
XJ11-1B12 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Images
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Western blot - Anti-Separase antibody [XJ11-1B12] (ab16170)Anti-Separase antibody [XJ11-1B12] (ab16170) at 10 µg/ml + MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 20 µg
Secondary
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
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![Immunocytochemistry/ Immunofluorescence - Anti-Separase antibody [XJ11-1B12] (ab16170)](https://www.abcam.com/ps/products/16/ab16170/Images/ab16170-235667-anti-separase-antibody-xj11-1b12-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Separase antibody [XJ11-1B12] (ab16170)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.
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![Flow Cytometry - Anti-Separase antibody [XJ11-1B12] (ab16170)](https://www.abcam.com/ps/products/16/ab16170/Images/ab16170-24881-anti-separase-antibody-xj11-1b12-flow-cytometry.jpg)
Flow Cytometry - Anti-Separase antibody [XJ11-1B12] (ab16170)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.
