CRISPR-Cas9 Antibody (1013816)

R&D Systems | Catalog # MAB10252

R&D Systems

Key Product Details

Species Reactivity

Multi-Species

Applications

Western Blot, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 1013816
View all CRISPR-Cas9 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived Cas9
Asp2-Asp1368
Accession # Q99ZW2

Specificity

Detects Cas9 in direct ELISAs and Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for CRISPR-Cas9 Antibody (1013816)

Detection of CRISPR-Cas9 antibody by Western Blot.

Detection of CRISPR-Cas9 by Western Blot.

Western blot shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with CRISPR-Cas9. PVDF membrane was probed with 2 µg/mL of Mouse Anti-CRISPR-Cas9 Monoclonal Antibody (Catalog # MAB10252) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for CRISPR-Cas9 at approximately 160 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human CRISPR-Cas9 antibody by Simple WesternTM.

Detection of Human CRISPR-Cas9 by Simple WesternTM.

Simple Western lane view shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with Cas9, loaded at 0.2 mg/mL. A specific band was detected for CRISPR-Cas9 at approximately 150 kDa (as indicated) using 20 µg/mL of Mouse Anti-CRISPR-Cas9 Monoclonal Antibody (Catalog # MAB10252). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for CRISPR-Cas9 Antibody (1013816)

Application
Recommended Usage

Simple Western

20 µg/mL
Sample: HEK293T human embryonic kidney cell line transfected with Cas9

Western Blot

2 µg/mL
Sample: HEK293T human embryonic kidney cell line transfected with CRISPR-Cas9

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CRISPR-Cas9

Streptococcus pyogenes Cas9 (CRISPR associated protein 9) is a 160 kDa RNA guided endonuclease that introduces site specific cleavage of double strand DNA (1). It is part of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system found in many bacteria such as S. pyogenes and most archaea, which provide adaptive immunity against invading mobile genetic elements (such as viruses, transposable elements and conjugative plasmids) (2, 3). Upon viral infection, short viral DNA (known as "spacers") integrate into the host genome between CRISPR repeats, and RNA sequences (guide RNA or gRNA) with this genetic information help guide Cas9 protein to recognize and cut foreign DNA. Cas9 protein undergoes conformational changes upon gRNA binding that shift from non-DNA binding conformation into an active DNA binding conformation. In the Cas9-gRNA complex, the gRNA sequence remains accessible to interact with free DNA, and the extent to which the gRNA spacer and target DNA segment (known as "protospacer") match will determine the cut site (4). The presence of a 5′-NGG-3′ protospacer adjacent motif (PAM) sequence immediately downstream of protospacers is required for Cas9 cleavage of the foreign DNA. PAM is absent in bacterial CRISPR loci, therefore preventing cleavage of the host genome (4). Cas9 associates with other proteins of the acquisition machinery (Cas1, Cas2 and Csn2), presumably to provide PAM specificity to this process (5). This RNA guided nuclease system called CRISPR/Cas (CRISPR associated protein) has been widely applied to genome engineering with increased efficiency (6). The attached nuclear localization signals(NLSs) on the chimeric protein ensures nuclear compartmentalization in mammalian cells during gene editing (7).

References

  1. Feng, Z. et al. (2013) Cell 154:1380.
  2. Moineau. et al. (2010) Nature 468:67.
  3. Barrangou, R. et al. (2014) Molecular Cell 54:234.
  4. Charpentier, E. et al. (2011) Nature 471:602.
  5. Leler, R. et al. (2015) Nature 519:199.
  6. Thomson, J.A. et al. (2013) PNAS,110:15644.
  7. Cong, L. et al. (2013) Science 339:819.

Long Name

CRISPR-associated Protein 9

Alternate Names

Cas9, CRISPR-associated endonuclease Cas9/Csn1, CRISPR-Cas9/Csn1, CRISPR/Cas9, csn1, SPy_1046, SPy1046, SpyCas9

Entrez Gene IDs

901176 (S. pyogenes)

Gene Symbol

CAS9

UniProt

Q99ZW2

Additional CRISPR-Cas9 Products

Product Documents for CRISPR-Cas9 Antibody (1013816)

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for CRISPR-Cas9 Antibody (1013816)

For research use only

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