| Description | EnzymoPure™Q M-MLV Reverse Transcriptase uses single-stranded RNA or DNA in the presence of a primer to synthesize a complementary DNA strand with high fidelity, sensitivity, and specificity. It is a modified, optimized reverse transcriptase particularly suitable for qPCR analysis. The EnzymoPure™Q M-MLV Reverse Transcriptase uses single-stranded RNA or DNA in the presence of a primer to synthesize a complementary DNA strand with high fidelity, sensitivity, and specificity. It is a modified, optimized reverse transcriptase particularly suitable for qPCR analysis. The EnzymoPure™Q M-MLV Reverse Transcriptase contains RNase H enzyme activity that can specifically degrade the RNA strand of an RNA-DNA hybrid, enabling the subsequent synthesis of second-strand cDNA.EnzymoPure™Q M-MLV Reverse Transcriptase, a commonly used high-quality reverse transcriptase, is widely used in the synthesis of first-strand cDNA using total RNA or mRNA templates, and is particularly suitable for qPCR and one-step qRT-PCR analysis. This product is also applicable for conventional PCR, synthesis of second-strand cDNA and construction of cDNA library, and cloning of target genes. It can also be used for DNA probe labeling with fluorescence, biotin, digoxin, or isotope through reverse transcription reaction, or for RNA studies by primer extension.Please refer to Figure 1 for the qPCR analysis of the GADPH gene in HEK293T cells using the cDNA templates obtained by the EnzymoPure™Q M-MLV reverse transcriptase.Figure 1Source：Recombinant reverse transcriptase expressed in E. coli.Definition of enzyme activity unit: One unit is defined as the amount of enzyme that incorporates 1 nmol of dTTP into acid-precipitable material in 10min at 37℃ using poly(A)•oligo(dT)12-18 as template-primer. Reaction system: 50mM Tris-HCl (pH8.3), 75mM KCl, 3mM MgCl2, 10mM DTT, 0.5mM [3H]-dTTP, and 0.4mM polyA•oligo(dT)12-18.Purity: This product is free from DNA endonuclease, exonuclease, phosphatase, and RNase, and can meet the requirements of conventional reverse transcription.Storage buffer: 20mM Tris-HCl (pH7.5), 150mM NaCl, 0.1mM EDTA, 1mM DTT, 0.01%(v/v) NP-40, and 50%(v/v) glycerolInactivation or inhibition：RT Q M-MLV Reverse Transcriptase can be inactivated by incubation at 80℃ for 10 minutes, or inhibited by EDTA and EGTA chelators, inorganic phosphates or pyrophosphates, and polyamine.The concentration of M-MLV is 200U/µl. When 20µl of reverse transcription reaction volume is used, D7188S, D7188M, and D7188L are sufficient for 50, 250, and 1000 reactions, respectively. provides a variety of reverse transcriptase. To select the best one for your needs, please refer to the following webpage:http://www.aladdin-e.com/support/reversetranscriptase.htmPrecautions：Please refer to the instructions for reverse transcription of RNAs with high GC content.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use：1. First-strand cDNA synthesisa. Set up the following reaction on ice or at room temperature. RNase Inhibitor and dNTP mix can be purchased from SYBR Green qPCR Mix (2X), SYBR Green qPCR Mix (2X, Low ROX), and SYBR Green qPCR Mix (2X, High ROX) are for conventional qPCR analysis with SYBR Green fluorescent dye. Probe qPCR Mix (2X), Probe qPCR Mix (2X, Low ROX), and Probe qPCR Mix (2X, High ROX) are for qPCR analysis with Taqman Probe.b. Perform qPCR analysis following the instructions of the corresponding qPCR kit.3. For other applications such as primer extension and probe labeling, please refer to references related to M-MLV reverse transcriptase.FAQ:1. The reverse transcription product of total RNA is invisible after electrophoresis.It is a normal phenomenon, because the amount of RNA template is low, and the amount of reverse transcription products in different sizes is even lower. 2. No specific product is amplified from the reverse transcription product.a. To exclude the problem of PCR reaction system or reverse transcription product, use gene-specific primers to amplify internal reference genes, such as actin and GAPDH. If reference genes can be amplified, but not the target gene, it indicates primers of the target gene are not well designed. b. Template RNA may be degraded. The integrity of total RNA can be checked by agarose gel or on-chip electrophoresis. Intact total RNA exhibits sharp, clear 28S and 18S rRNA bands, and the 28S rRNA band should be approximately twice as intense as the 18S rRNA band. A ratio less than 2 indicates the degradation of total RNA and new total RNA should be prepared. c. RNA samples may contain some components that inhibit the activity of reverse transcriptase. Those contaminants include phenol, SDS, EDTA, guanidine salts, phosphoric acid, pyrophosphoric acid, polyamine, spermidine, etc, which can be removed effectively by column purification or precipitation, washing, and redissolution. Total RNA extracted by Zol or Trizol produced by can fully meet the requirements of reverse transcription.d. Insufficient templates for reverse transcription. When DNase I is used to remove the residual DNA in the RNA sample prior to reverse transcription and is subjected to heat-inactivation, EDTA should be added to the RNA sample at a final concentration of 2.5mM to protect RNA from degradation under high temperature. Additionally, to amplify a specific gene, it is necessary to extract RNA from tissues in which the target gene is highly expressed. e. An inappropriate primer is used for reverse transcription. Random hexamer instead of Oligo(dT)18 should be used for the reverse transcription of bacterial total RNA which does not have poly(A) tails. Gene-specific primers used for reverse transcription must be well-designed... Read More | The content of this cell is too long for an XLSX file (more than 32767 characters). Please use the CSV format for this export | C1q separated from C1r and C1s and from other stabilizing proteins tends to aggregate easily. Because it was isolated and studied in numerous research laboratories, many buffers have been used to stabilize concentrated C1q and prevent aggregation. About half of the scientists prefer high salt and C1q separated from C1r and C1s and from other stabilizing proteins tends to aggregate easily. Because it was isolated and studied in numerous research laboratories, many buffers have been used to stabilize concentrated C1q and prevent aggregation. About half of the scientists prefer high salt and the other prefer 40% glycerol in the storage buffer.C1q is purified from pooled normal human plasma. C1q is part of the C1 complex and this complex is the first complement component in the cascade referred to as the classical pathway of complement. C1 is actually a non-covalent assembly of three different proteins (C1q, C1r, and C1s) bound together in a calcium-dependent complex. C1q has six extended arms with domains at the end of each arm that bind to the Fc domains of immunoglobulins. When antibodies bind to antigens forming immune complexes they cluster allowing two or more of its six arms of C1q to bind to the Fc domains of antibodies such as IgG or IgM. The binding of multiple arms to immune complexes causes the two C1r proteins in the complex (protease zymogens) to auto-activate producing two C1r proteases that cleave and activate the two C1s protease zymogens in the complex. Activated C1s cleaves complement component C4 releasing C4a and initiating covalent attachment of C4b to the activating surface. Activated C1s also cleaves C2 and the larger fragment of C2 binds to the surface-attached C4b forming C4b,C2a which is the C3/C5 convertase of the classical pathway.Extinction Coeff.A₂₈₀ nm = 0.68 at 1.0 mg/ml for pure C1q Molecular weight:410,000 Da (18 chains)Preservative:None, 0.22 µm filtered.Source:Normal human serum (shown by certified tests to be negative for HBsAg, HTLV-I/II, STS, and for antibodies to HCV, HIV-1 and HIV-II).Physical Characteristics & StructureC1q is a high molecular weight complex of 18 polypeptide chains. Each of the six arms of C1q contains three chains, an A chain (26,000 daltons), a B chain (25,000 daltons) and a C chain (24,000 daltons). The three chains are coiled into a collagen-like triple helix over approximately half their length. Half of this collagen region forms a central core where all 18 chains come together. The chains are joined in this core by disulfides in the pattern A-B and C-C. There is a bend in the center of the collagen region allowing the arms to extend away from each other. Globular heads at the far ends of the collagen arms possess binding sites for Fc domains of immunoglobulins. C1 complex is composed of one C1q molecule (410,000 daltons), two C1r molecules (92,000 daltons) and two C1s molecules (86,000 daltons). The complex is stable in the presence of calcium, but easily dissociates if calcium is removed. When C1 is activated the C1r and C1s subunits are each cleaved into two chain molecules due to proteolytic activation. Thus, the SDS gel pattern of C1 is very complex. Function The biological functions of C1q are described above in the General Description and Physical Characteristics sections. C1q functional activity may be assayed using C1q-depleted serum and EA cells. These assays are extremely sensitive to C1q typically yielding 50% lysis with less than 2 ng C1q in assays measuring the lysis of EA cells. AssaysThe unit of classical pathway activity is the CH50. A similar unit, the C1qH50, is used to quantitate the activity of C1q. A C1qH50 unit is the amount of functional C1q needed to lyse 50% of 3×10^7 EA cells (antibody-sensitized sheep erythrocytes) when that amount of C1q is incubated with 5-20 µL of C1q-Dpl in GVB++ in a total volume of 500 µL for 30 min at 37℃. This amount of C1q indicates the sensitivity of the assay for C1q which is typically about 1 ng C1q with 10 µL C1q-Dpl. See the Certificate of Analysis for lot specific values.ApplicationsC1q is used to coat ELISA plates to capture and quantitate immune complexes in clinical samples. A number of commercial companies sell diagnostic kits for immune complex detection and quantitation. These kits are based on the ability of C1q to bind well to immune complexes, but to not bind significantly to monomeric immunoglobulins. GeneticsThe EMBL/Genbank cDNA accession numbers are: C1q A chain (P02745), C1q B chain (P02746), and C1q C chain (P02747). The genes for C1q chains A, B and C are all located on chromosome 1p in the order A-C-B. DeficienciesDeficiencies of each of the three components of C1 have been found. Patients lacking C1q generally have immune-complex-mediated renal disease and skin lesions. Like all patients lacking early classical pathway components C1q deficient individuals are prone to systemic lupus erythrematosis (SLE) and recurrent pyogenic infections. They lack classical pathway function and may or may not exhibit C1q antigen in blood.DiseasesSee section titled Deficiencies above. Precautions/Toxicity/HazardsThis protein is purified from human serum and therefore precautions appropriate for handling any blood-derived product must be used even though the source was shown by certified tests to be negative for HBsAg, HTLV-I/II, STS, and for antibodies to HCV, HIV-1 and HIV-II... Read More | Collagenase NB 1 is chromatographically highly purified; therefore it contains a very high collagenolytic activity. It is largely free from additional enzymatic activities like clostripain, trypsin-like activity and neutral protease, as well as endotoxins.SpecificationsContains chromatographically Collagenase NB 1 is chromatographically highly purified; therefore it contains a very high collagenolytic activity. It is largely free from additional enzymatic activities like clostripain, trypsin-like activity and neutral protease, as well as endotoxins.SpecificationsContains chromatographically highly purified class I and class II collagenase (1).Largely free from clostripain, trypsin-like protease and neutral protease.Vial contains not less than 2000 PZ U collagenase.Activity (Wünsch): ≥ 3.00 U/mgEndotoxin: ≤ 10.0 EU/mg (Ph. Eur.)(Clostridiopeptidase A)EC 3.4.24.3 • Mr ca. 70 000 - 120 000 (collagenases) • CAS [9001-12-1]ApplicationCollagenase NB 1 is, mostly in combination with Neutral Protease NB, suitable for cell isolation from several tissue types.References and DefinitionsUnit definition: Collagenase: 1 U according to Wünsch (2) catalyzes the hydrolysis of 1 µmole 4-phenylazobenzyloxycarbonyl-L-prolyl-L-leucylglycyl-L-prolyl-D-arginine per minute at 25 °C, pH 7.1.Endotoxin: Ph. Eur. (1 Endotoxin Unit is equal to 1 International Unit of a WHO approved reference standard endotoxin (RSE)).References1. Bond, M.D. & van Wart, H.E. (1984) Biochemistry 23, 3077-30912. Wünsch, E. & Heidrich, H.G. (1963) Hoppe-Seyler's Z. Physiol. Chem. 333, 149-51... Read More | Inquire |