Revision 3

#50574

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, W-S, IP

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
233, 160

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q14674

Entrez-Gene Id:
9700

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

Separase (F4R3M) Rabbit Monoclonal Antibody recognizes endogenous levels of total separase protein. This antibody is expected to detect full-length isoform 1 and the N-terminal cleavage fragment. This antibody is not expected to detect the C-terminal cleavage fragment. Based on the peptide antigen sequence, this antibody is expected to recognize isoform 2. This antibody detects an approximately 40 kDa protein of unknown identity in some cell lines.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg1320 of human separase protein.

Background

Caspase-like protein ESPL1 (separase), also known as separin, is a protease critical for chromosome segregation. It plays a role in spindle stabilization and elongation, apoptosis, and DNA repair. Separase cleaves the SCC1/RAD21 subunit of the cohesin complex at the start of anaphase, allowing sister chromatid segregation, an irreversible step in cell division (1). Separase is negatively regulated by securin, which inhibits its proteolytic activity, and by CDK1/cyclin B-dependent phosphorylation. Both securin and cyclin B are degraded via the anaphase-promoting complex/cyclosome (APC/C) (2,3). Dysregulation of separase activity is linked to cancer and genome instability, making it a target for cancer drug discovery (4,5). Separase is overexpressed in human cancer, and overexpression is thought to induce aneuploidy and tumorigenesis (1,6).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation

Cross-Reactivity Key

H: Human

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Western blot analysis of extracts from NCI-H1299 cells, transfected with control siRNA (-) or human Separase siRNA (hSeparase siRNA; +), using Separase (F4R3M) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 1: Separase (F4R3M) Rabbit Monoclonal Antibody
Western blot analysis of extracts from various cell lines using Separase (F4R3M) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of separase protein in RL and Caki-1 cells is consistent with the predicted expression pattern.
Western Blotting Image 2: Separase (F4R3M) Rabbit Monoclonal Antibody
Simple WesternTM analysis of lysates (0.1 mg/mL) from NCI-H1299 using Separase (F4R3M) Rabbit mAb #50574. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66 - 440 kDa separation module.
Western Blotting Image 1: Separase (F4R3M) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Immunoprecipitation of separase protein from NCI-H1299 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Separase (F4R3M) Rabbit mAb. Western blot analysis was performed using Separase (F4R3M) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: Separase (F4R3M) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.