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73211
Cas9 and Associated Proteins Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Cas9 and Associated Proteins Antibody Sampler Kit #73211

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Confocal immunofluorescent analysis of fixed frozen brain from constitutive Cas9 (S. pyogenes)-expressing mice (left, positive) or wild type mice (right, negative), using Cas9 (S. pyogenes) (D8Y4K) Rabbit mAb (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing AsCpf1/Cas12a (Strain BV3L6), FnCpf1/Cas12a (Strain U112), Cas9 (S. pyogenes), or Cas9 (S. aureus) (+), using AsCpf1/Cas12a (Strain BV3L6) (E1U7C) Rabbit mAb (upper) and Myc-Tag (71D10) Rabbit mAb #2278 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Cas9 (S. aureus), Cas9 (S. pyogenes), AsCpf1 (Strain BV3L6), or FnCpf1 (Strain U112) (+), using Cas9 (S. aureus) (E4G3U) Rabbit mAb (upper) and Myc-Tag (71D10) Rabbit mAb #2278 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Cas9 (S. pyogenes) (+), using Cas9 (S. pyogenes) Rabbit mAb mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing FnCpf1/Cas12a (Strain U112), AsCpf1/Cas12a (Strain BV3L6), Cas9 (S. pyogenes), or Cas9 (S. aureus) (+), using FnCpf1/Cas12a (Strain U112) (E7I2B) Rabbit mAb (upper) and Myc-Tag (71D10) Rabbit mAb #2278 (lower).
Confocal immunofluorescent analysis of 293T cells transfected with a Myc-tagged AsCpf1/Cas12a (Strain BV3L6) construct using AsCpf1/Cas12a (Strain BV3L6) (E1U7C) Rabbit mAb (green) and Myc-Tag (9B11) Mouse mAb #2276 (red). Overlapping signals are shown in yellow. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of 293T cells transfected with a construct expressing myc-tagged Cas9 (S. aureus) using Cas9 (S. aureus) (E4G3U) Rabbit mAb (green) and Myc-Tag (9B11) Mouse mAb #2276 (red). Overlapping signals are shown in yellow. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of Cas9-expressing 293T cells (left) and unmodified 293T cells (right) using Cas9 (S. pyogenes) (D8Y4K) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).
Confocal immunofluorescent analysis of 293T cells transfected with a Myc-tagged FnCpf1/Cas12a (Strain U112) construct using FnCpf1/Cas12a (Strain U112) (E7I2B) Rabbit mAb (green) and Myc-Tag (9B11) Mouse mAb #2276 (red). Overlapping signals are shown in yellow. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of 293 cells, untransfected (blue) or transfected with myc-tagged AsCpf1/Cas12a (Strain BV3L6) (green), using AsCpf1/Cas12a (Strain BV3L6) (E1U7C) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow cytometric analysis of 293T cells, untransfected (blue) or transfected with a construct expressing myc-tagged Cas9 (S. aureus) (green), using Cas9 (S. aureus) Rabbit mAb (solid line) compared to a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow cytometric analysis of 293T cells, untreated (blue) or transfected with S. pyogenes plasmid (green), using Cas9 (S. pyogenes) (D8Y4K) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow cytometric analysis of 293 cells, untransfected (blue) or transfected with a construct expressing myc-tagged FnCpf1/Cas12a (Strain U112) (green), using FnCpf1/Cas12a (Strain U112) (E7I2B) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
To Purchase # 73211
Cat. # Size Qty. Price
73211T
1 Kit  (4 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Cas9 (S. pyogenes) (D8Y4K) Rabbit mAb 65832 20 µl
  • WB
  • IF
  • F
All 150 Rabbit IgG
Cas9 (S. aureus) (E4G3U) Rabbit mAb 51610 20 µl
  • WB
  • IF
  • F
All 124 Rabbit IgG
AsCpf1/Cas12a (Strain BV3L6) (E1U7C) Rabbit mAb 19984 20 µl
  • WB
  • IF
  • F
All 151 Rabbit IgG
FnCpf1/Cas12a (Strain U112) (E7I2B) Rabbit mAb 90111 20 µl
  • WB
  • IF
  • F
All 152 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Rab Goat 

Product Description

The Cas9 and Associated Proteins Antibody Sampler Kit provides an economical means of detecting Cas9 and Cas9-related family members. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Cas9 and Associated Proteins Antibody Sampler Kit recognizes transfected levels of its target protein. Antibodies are specific for the indicated endonuclease target and cross-reactivity with other endonucleases is not observed.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Val16 of Cas9 (S. pyogenes), Val905 of Cas9 (S. aureus), Leu822 of Acidaminococcus sp. Cpf1 (Strain BV3L6), and Ile841 of Cpf1 from Francisella tularensis subsp. novicida (Strain U112).

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). Class 2 CRISPR systems rely on single-component effector proteins to mediate DNA interference (2). Several Class 2 CRISPR effector proteins, derived from specific bacterial species, are used for genome editing. Cas9 family of proteins, derived from S. pyogenes and S. aureus, are some of the most well characterized and widely used editing effector enzymes. Additional members of the Class2 CRISPR system include Cpf1/Cas12a (CRISPR from Prevotella and Francisella) endonucleases (3). Cpf1/Cas12a endonucleases, compared to Cas9 systems, have several unique features that increase the utility of CRISPR-based genome editing techniques: 1) Cpf1/Cas12a-mediated cleavage relies on a single and short CRISPR RNA (crRNA) without the requirement of a trans-activating crRNA (tracrRNA), 2) Cpf1/Cas12a utilizes T-Rich protospacer adjacent motif (PAM) sequences rather than a G-Rich PAM, and 3) Cpf1/Cas12a generates a staggered, rather than a blunt-ended, DNA double-stranded break (3). These features broaden the utility of using CRISPR-Cas systems for specific gene regulation and therapeutic applications. Several Cpf1/Cas12a bacterial orthologs, e.g. Francisella novicida U112 and Acidaminococcus sp. BV3L6, have been characterized for CRISPR-mediated mammalian genome editing (3,4).

Limited Uses

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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.

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