Product Number: RPA008

Shipping and Storage

Store at -20℃ for two years (it is recommended to store separately to avoid repeated freezing and thawing affecting protein activity).

Description

Creatine kinase (CK), also known as creatine kinase or phosphocreatine kinase. Creatine kinase is mainly present in skeletal muscle, myocardium, and smooth muscle, followed by brain tissue, with lower levels in the gastrointestinal tract, lungs, and kidneys. Creatine kinase mainly exists in the cytoplasm and mitochondria, and is an important kinase directly related to intracellular energy transport, muscle contraction, and ATP regeneration. Creatine kinase activity assay can be used for the diagnosis of skeletal muscle diseases and myocardial diseases.

Our company provides genetically recombinant creatine kinase (CK, or CK, muscle, CKM) derived from animal muscles, with high activity and good stability.

Application

DNA isothermal amplification; RPA amplification; Genetic testing.

Specification

1. Dosage form: liquid or freeze-dried powder
2. Storage buffer: 20mM Tris, pH 8.0
3. Enzyme activity definition: At 37℃, one unit of enzyme can transfer 1.0 micromolar phosphate from phosphocreatine to ADP per minute (measured at 340nm, based on the coupling reaction producing 1 equimolar amount of NADH).
4. Source: Recombinant gene expression
5. Molecular weight: around 43KDa (detected by SDS-PAGE)
6. Purity: ≥ 90% (detected by SDS-PAGE)

Note

Not suitable for human experiments.

Product Number: RPA007-100

Shipping and Storage

Store at -20℃ for two years (it is recommended to store separately to avoid repeated freezing and thawing affecting protein activity).

Description

Bsu DNA polymerase is a DNA isothermal amplification polymerase with chain displacement activity, mainly used for RPA recombinase amplification. The protein DNA complex formed by the combination of recombinant enzyme UvsX and primers can search for homologous sequences in double stranded DNA; Once the primer locates the homologous sequence, a chain exchange reaction occurs, and Bsu DNA polymerase initiates DNA synthesis, exponentially amplifying the target region on the template. The replaced DNA strand binds to SSB to prevent further substitution.

Our company provides Bsu DNA polymerase for gene recombinant expression, with a molecular weight of~68kDa, high activity, and good stability. This product is a large fragment of Bsu DNA polymerase, derived from thermophilic Bacillus subtilis. It is obtained by truncating the first 296 AA of the Bacillus subtilis DNA polymerase (Bsu) I gene. This enzyme retains the 5 ‘-3’ polymerase activity of Bsu I, but lacks the 5 ‘-3’ exonuclease domain. The large fragment itself lacks 3 ‘-5’ exonuclease activity and can be used for recombinant enzyme amplification.

We constructed an efficient RPA isothermal amplification system using self-produced large fragments of Bsu DNA polymerase (as shown in the figure below).

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A RPA isothermal amplification system was constructed using self-produced recombinant proteins such as Uxsx, UxsX, gp32, Bsu, etc.

M is a marker, and 1, 2, 3, and 4 are different RPA constant temperature amplification products.

Application

RPA isothermal amplification; RPA chain substitution for DNA synthesis; Random primer labeling method; Synthesis of the second strand of cDNA; The suffix of a single dA.

Specification

1. Dosage form: liquid or freeze-dried powder
2. Storage buffer: 50mM Tris HCl, 50mM KCl, 1mM DTT, 0.1mM EDTA, 20% glycerol, pH 7.5
3. Molecular weight: around 68 KDa (detected by SDS-PAGE)
4. Purity: ≥ 90% (detected by SDS-PAGE)
5. Thermal deactivation: 75℃, 20min

Note

1. Due to the lack of 3 “-5” exonuclease activity, the large fragment of Bsu DNA polymerase cannot cleave the 3 “unpaired protruding end, making it unsuitable for generating a flat end.
2. At 25 ℃, the large fragment of Bsu DNA polymerase retains 50% of its activity, which is twice that of the Klenow fragment (3 “-5” exo -) at the same temperature.
3. This reagent is only used for research and development or production, and is strictly prohibited from being used in human or animal experiments.

Product Number: RPA002

Shipping and Storage

Store at -20℃ to avoid repeated freezing and thawing.

Description

UvsY is a bacteriophage T4 recombinant regulatory protein that plays a promoting role in the homologous recombination process of UvsX. When searching for homologous sequences in UvsX, it needs to compete for binding sites with single stranded DNA binding proteins pre bound to single stranded DNA, and UvsY protein promotes this competition, which helps UvsX bind to single stranded DNA. It can enhance the DNA dependent ATPase activity of UvsX protein, reduce the minimum concentration required for its activity, and promote chain replacement. UvsY promotes the invasion of UvsX recombinase into ssDNA single stranded binding protein complex, leading to the release of single stranded binding protein, thereby promoting the binding of UvsX to single stranded DNA. This enzyme has no nuclease activity.

Our company provides gene recombinant expression of UvsY protein, with a molecular weight of~16KDa, high activity, and good stability. We constructed an efficient RPA isothermal amplification system using self-produced UvsY protein (as shown in the figure below).

A RPA isothermal amplification system was constructed using self-produced recombinant proteins such as Uxsx, UxsX, gp32, Bsu, etc.

M is a marker, and 1, 2, 3, and 4 are different RPA constant temperature amplification products.

Application

DNA isothermal amplification; RPA amplification; Genetic testing.

Specification

1. Dosage form: liquid or freeze-dried powder
2. Storage buffer: 50mM Tris HCl, 50mM KCl, 1mM DTT, 0.1mM EDTA, 20% glycerol, pH 8.0
3. Molecular weight: Approximately 16 KDa (detected by SDS-PAGE)
4. Purity: ≥ 90% (detected by SDS-PAGE)
5. Thermal deactivation: 60℃, 10 minutes

Quality control

1. Detection of residual nucleases: Incubate 500U of this enzyme with 0.6μg of λ – Hind III at 37℃ for 16 hours, and the electrophoresis band of DNA remains unchanged.
2. Detection of residual endonuclease: 500U of this enzyme and 0.6μg Supercoiled pBR322 DNA were incubated at 37℃ for 4 hours, and the electrophoresis band of DNA remained unchanged.
3. E. coli DNA residue detection: The residual nucleic acid in 50U of this product was detected by TaqMan qPCR specific for E. coli 16s rDNA, and the E. coli genome residue was less than 10 copies.

Note

This reagent is only used for research and development or production, and is strictly prohibited from being used in human or animal experiments.

Product Number: RPA001

Shipping and Storage

Store at -20℃ to avoid repeated freezing and thawing.

Description

Recombinant enzyme polymerase amplification technology is a new technology that involves multiple enzymes and proteins to achieve nucleic acid index amplification under constant temperature conditions. It has the characteristics of sensitive reaction, high efficiency, and high cost-effectiveness. Recombinant enzyme polymerase amplification technology simulates DNA amplification in vivo and produces the target fragment under isothermal conditions. This technology mainly relies on the recombinase UvsX, single stranded binding protein gp32, and strand displacement DNA polymerase Bsu. The recombinase UvsX is capable of breaking double stranded DNA without heating. At the beginning of the recombinase polymerase amplification reaction, the recombinase UvsX binds to the primer with the participation of ATP, forming nucleic acid protein complexes that can scan for target double stranded DNA complementary to the primer sequence. Subsequently, the nucleic acid protein complex invades the 5 ‘end site to form a D-shaped loop. The single chain binding protein gp32 binds to the displaced single chain to stabilize it. At the same time, the recombinant enzyme uvsx leaves the 3 ‘end of the oligonucleotide and is degraded before being utilized by DNA polymerase. Afterwards, the strand displacement DNA polymerase Bsu binds to the free 3 ‘end of the nucleic acid protein complex for strand extension, forming a new complementary strand. During the process of chain extension, the newly synthesized single chain pairs with the original complementary chain. The above steps are repeated to achieve exponential growth of DNA.

Our company provides recombinant expression of UvsX protein with a molecular weight of~44kDa, high activity, and good stability. We constructed an efficient RPA isothermal amplification system using self-produced UvsX protein (as shown in the figure below).

A RPA isothermal amplification system was constructed using self-produced recombinant proteins such as Uxsx, UxsX, gp32, Bsu, etc.

M is a marker, and 1, 2, 3, and 4 are different RPA constant temperature amplification products.

Application

DNA isothermal amplification; RPA amplification; Genetic testing.

Specification

1. Dosage form: liquid or freeze-dried powder
2. Storage buffer: 50mM Tris HCl, 50mM KCl, 1mM DTT, 0.1mM EDTA, pH 8.0
3. Molecular weight: Approximately 44 KDa (detected by SDS-PAGE)
4. Purity: ≥ 90% (detected by SDS-PAGE)
5. Thermal deactivation: 60℃, 10 minutes

Quality control

1. Detection of residual nucleases: Incubate 500U of this enzyme with 0.6μg of λ – Hind III at 37℃ for 16 hours, and the electrophoresis band of DNA remains unchanged.
2. Detection of residual endonuclease: 500U of this enzyme and 0.6μg Supercoiled pBR322 DNA were incubated at 37℃ for 4 hours, and the electrophoresis band of DNA remained unchanged.
3. E. coli DNA residue detection: The residual nucleic acid in 50 U of this product was detected by TaqMan qPCR specific for E. coli 16s rDNA, and the E. coli genome residue was less than 10 copies.

Note

This reagent is only used for research and development or production, and is strictly prohibited from being used in human or animal experiments.

Product Number: RPA006

Shipping and Storage

1. Storage conditions: Store at -20℃ to avoid repeated freezing and thawing.
2. Transportation conditions: Low temperature ice packs.

Description

Gp32 protein (ssDNA binding protein), also known as T4 gp32 protein or T4 phage gp32 protein, is a single stranded DNA (ssDNA) binding protein encoded by gene 32 of the T4 phage genome. This protein is essential for T4 phage DNA replication and repair, and can be used for recombinase polymerase amplification (RPA). When observing the DNA structure inside cells using an electron microscope, this protein is also widely used to stabilize and label single stranded DNA regions. In addition, T4 gp32 has been reported to promote restriction endonuclease and T4 DNA polymerase activity, improve reverse transcriptase efficiency during RT-PCR, and increase PCR product yield.

Our company provides recombinant expression of gp32 protein with a molecular weight of~35 KDa, high activity, and good stability. We constructed an efficient RPA isothermal amplification system using self-produced gp32 protein (as shown in the figure below).

Application

Stabilize or label ssDNA; Increase the production of reverse transcription products during RT-PCR process; Enhance the synthesis of DNA in vitro; Used together with T4 DNA polymerase and T4 DNA ligase for site-specific mutation experiments; When amplifying bacterial or fungal genomes from samples containing humic acid, such as soil, the yield and specificity of PCR products can be increased; Promote the digestion reaction of restriction endonucleases.