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  3. CRISPR-Cas9 Protein, S. pyogenes (His)

CRISPR-Cas9 Protein, S. pyogenes (His)

Cat. No.: HY-P77727
COA Handling Instructions

The CRISPR-Cas9 protein is part of an immune system that defends against genetic elements. It processes RNA and aids in cleaving DNA targets. Protein and guide RNAs are necessary for its function. The protein recognizes specific sequences to distinguish self from nonself and provides immunity against matching genetic elements. CRISPR-Cas9 Protein, S. pyogenes (His) is the recombinant CRISPR-Cas9 protein, expressed by E. coli , with His tagged. The total length of CRISPR-Cas9 Protein, S. pyogenes (His) is 1367 a.a., with molecular weight of ~163 kDa.

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  • Biological Activity

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Description

The CRISPR-Cas9 protein is part of an immune system that defends against genetic elements. It processes RNA and aids in cleaving DNA targets. Protein and guide RNAs are necessary for its function. The protein recognizes specific sequences to distinguish self from nonself and provides immunity against matching genetic elements. CRISPR-Cas9 Protein, S. pyogenes (His) is the recombinant CRISPR-Cas9 protein, expressed by E. coli , with His tagged. The total length of CRISPR-Cas9 Protein, S. pyogenes (His) is 1367 a.a., with molecular weight of ~163 kDa.

Background

CRISPR-Cas9, a revolutionary molecular tool, constitutes an adaptive immune system that defends against mobile genetic elements like viruses, transposable elements, and conjugative plasmids. In this system, CRISPR clusters contain spacers, which are sequences complementary to preceding mobile elements, serving as a memory bank of past encounters. The process involves transcription and processing of CRISPR RNA (crRNA), wherein, for type II CRISPR systems, the proper processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc), and the pivotal Cas9 protein. The tracrRNA guides ribonuclease 3 in processing pre-crRNA, with Cas9 stabilizing the pre-crRNA:tracrRNA interaction. In subsequent steps, Cas9/crRNA/tracrRNA complex acts as an endonuclease, cleaving the target DNA with complementary spacer sequences. Cas9 is inactive without the guidance of two specific guide RNAs (gRNA), emphasizing the necessity of both gRNAs for DNA binding and nuclease activity. Cas9's recognition of the protospacer adjacent motif (PAM) within CRISPR repeats is crucial for distinguishing self from nonself, ensuring accurate target identification. This sophisticated system confers immunity against invading genetic elements through a mechanism known as CRISPR interference.

Biological Activity

The cleavage activity in vitro was not less than 85%.

Species

Others

Source

E. coli

Tag

His

Accession

Q99ZW2 (D2-D1368)

Gene ID

/

Molecular Construction
N-term
CRISPR-Cas9 (D2-D1368)
Accession # Q99ZW2
C-term
Synonyms
CRISPR-associated endonuclease Cas9/Csn1; cas9; SpCas9; SpyCas9
Molecular Weight

Approximately 163 kDa

Purity

Greater than 95% as determined by Tris-Bis PAGE.

Appearance

Solution.

Formulation

Supplied as 0.22 μm filtered solution in 30 mM Tris-HCl, 0.3 M NaCl, 50% glycerol, 0.1 mM EDTA, pH 7.4.

Endotoxin Level

<0.01 EU/μg, determined by LAL method.

Reconstitution

N/A.

Storage & Stability

Stored at -80°C for 1 year. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping

Shipping with dry ice.

Documentation

CRISPR-Cas9 Protein, S. pyogenes (His) Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

  • Reconstitution Calculator

  • Dilution Calculator

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The reconstitution calculator equation

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration
= ÷

The dilution calculator equation

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
× = ×
C1   V1   C2   V2

The specific activity calculator equation

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)
Unit/mg = 106 ÷ ng/mL

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Product Name:
CRISPR-Cas9 Protein, S. pyogenes (His)
Cat. No.:
HY-P77727
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