Revision 4

#53413Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, W-S

REACTIVITY:

All

SENSITIVITY:

Transfected Only

MW (kDa):

70

Source/Isotype:

Mouse IgG1

UniProt ID:

#A0A482D308

Product Information

Product Usage Information

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

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

CasMINI (8H6-A8) Mouse mAb recognizes transfected levels of CasMINI protein. This antibody may detect several non-specific bands of unknown origin in rat cell lines.

Species Reactivity:

All Species Expected

Source / Purification

Monoclonal antibody is produced by immunizing animals with full-length recombinant protein specific to CasMINI protein.

Background

CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins) are RNA-guided nuclease effectors that are utilized for precise genome editing in mammalian systems (1). Cpf1/Cas12a (CRISPR from Prevotella and Francisella) proteins are members of the Class 2 CRISPR system (2). Class 2 CRISPR systems, such as the well characterized Cas9, rely on single-component effector proteins to mediate DNA interference (3). Cpf1/Cas12a and Class 2 CRISPR endonucleases permit efficient and specific genome editing that may be used as gene therapies for genetic diseases. However, virus-based vehicles used for in vivo delivery, like adeno-associated virus (AAV)-based vectors, have a limited packaging capacity. CasMINI was engineered from the naturally occurring type V-F Cas12f (Cas14) system. CasMINI is 529 amino acids, which is significantly smaller than Cpf1/Cas12a and Cas9 proteins (62% and 57% smaller, respectively), making CasMINI more suitable for virus-based packaging for in vivo delivery (4).

  1. Cong, L. et al. (2013) Science 339, 819-23.
  2. Zetsche, B. et al. (2015) Cell 163, 759-71.
  3. Horvath, P. and Barrangou, R. (2010) Science 327, 167-70.
  4. Xu, X. et al. (2021) Mol Cell 81, 4333-4345.e4.

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

WB: Western Blotting W-S: Simple Western™

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

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.

Revision 4
#53413

CasMINI (8H6-A8) Mouse mAb

Western Blotting Image 1: CasMINI (8H6-A8) Mouse mAb Expand Image
Western blot analysis of extracts from 293T cells, untransfected (-) or transfected with an amino-terminal CasMINI fusion protein (CasMINI-Myc; +) or a carboxy-terminal CasMINI fusion protein (Myc-CasMINI; +), using CasMINI (8H6-A8) Mouse mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: CasMINI (8H6-A8) Mouse mAb Expand Image
Simple WesternTM analysis of lysates (0.06 mg/mL) from 293T + CasMINI-Myc cells using CasMINI (8H6-A8) Mouse mAb #53414. 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 12-230 kDa separation module.