| Description | The Anstart Taq-D qPCR Master Mix (2×, with UDG) is a ready-to-use, probe-based qPCR master mix designed for sensitive and contamination-resistant fluorescence quantitative PCR detection in single or multiplex DNA targets. This product provides a simple, rapid, and highly sensitive PCR The Anstart Taq-D qPCR Master Mix (2×, with UDG) is a ready-to-use, probe-based qPCR master mix designed for sensitive and contamination-resistant fluorescence quantitative PCR detection in single or multiplex DNA targets. This product provides a simple, rapid, and highly sensitive PCR detection system, enabling real-time fluorescence PCR amplification in 35–40 minutes.Core ComponentsThe master mix contains:dNTP Mix (including dATP, dTTP, dCTP, dGTP, and dUTP)Antibody-modified hot-start DNA polymerase (Anstart Taq-D)UDG (Uracil-DNA Glycosylase)Users only need to add primers, probes (or fluorescent dyes), and template DNA to perform fluorescence-based qPCR.UDG Contamination Control MechanismDuring the initial 50°C, 2 min incubation, UDG cleaves the N-glycosidic bond between uracil and the sugar-phosphate backbone in any contaminating PCR products containing dUTP, releasing free uracil.Subsequent 95°C, 2.5 min heat treatment inactivates UDG while further degrading the phosphodiester backbone, effectively eliminating carryover contamination from uracil-containing PCR products.UDG acts on both single- and double-stranded DNA but is inactive against RNA.Compatible InstrumentsABI 7500, Qiagen QiaQuant 5-plexFor other instruments not listed, this kit has not been fully validated. If required, please contact our technical support team for further assistance.Protocol1. Reagent Preparation (Reagent Setup Area)1.1 Thaw the master mix at -20 ± 5°C, equilibrate to room temperature, vortex briefly, and centrifuge at low speed for 15 sec before use.Recommendation: Aliquot PCR reagents to avoid repeated freeze-thaw cycles.1.2 Prepare the PCR reaction mix according to the table below:ComponentVolume/25µLVolume/50µL2× Anstart Taq-D qPCR Master Mix12.5 µL25 µLPrimer-probe Mix1 µL2 µLTemplates5 µL10 µLddH2OUp to 25 µLUp to 50 µLTotal25 µL50 µLNote 1: Primer and template volumes can be adjusted as needed.Note 2: For fast cycling programs, a 25 µL reaction volume is recommended.1.3 Mix gently by pipetting or brief vortexing, then centrifuge briefly to collect liquid at the tube bottom.1.4 Transfer PCR tubes to the thermal cycler and initiate the reaction.2. Sample Preparation (Sample Prep Area)Add the appropriate sample volume as required.3. PCR Amplification (Amplification Area)3.1 Preheat the instrument and verify performance.3.2 Load prepared PCR tubes into the instrument and record tube positions. Set fluorescence channels as needed.Recommended Cycling ProgramsStandard ProgramStepCyclesTemp.TimeData Acquisition1150℃2minNo2195℃2min 30sNo34594℃10sNo44555℃40sYes5125℃10sNoFast ProgramStepCyclesTemp.TimeData Acquisition1150℃2minNo2195℃30sNo34594℃5sNo44555℃15sYes5125℃10sNoFluorescence acquisition can be set for 2-step or 3-step PCR depending on experimental needs.Optimal primer concentration: Typically 0.2 µM (Tm ≥ 55°C).Adjust annealing temperature and primer concentration for optimal amplification efficiency... Read More | 4-Methylumbelliferyl α-L-iduronide (free acid) is a fluorogenic substrate for α-L-iduronidase. This is found in cell lysosomes, which is involved in the degradation of glycosaminoglycans. 4-Methylumbelliferyl-α-L-iduronide is cleaved by α-L-iduronidase to release the fluorescent 4-Methylumbelliferyl α-L-iduronide (free acid) is a fluorogenic substrate for α-L-iduronidase. This is found in cell lysosomes, which is involved in the degradation of glycosaminoglycans. 4-Methylumbelliferyl-α-L-iduronide is cleaved by α-L-iduronidase to release the fluorescent moiety 4-methylumbelliferyl (4-MU). This 4-Methylumbelliferyl α-L-iduronide form is the free acid, which offers a considerable weight for weight advantage over the 4-MU iduronide salt in terms of its application dose.:For further studies, use α-L-iduronidase gene silencing:siRNA and shRNA:reagents and α-L-iduronidase gene editing:CRISPR:knockout and activation products... Read More | MAP kinase substrate (MBP) is a peptide substrate for ERK 1 and ERK 2 MAP kinases. Sequence contains amino acids 95-98 of myelin basic protein (MBP) including Thr 97 within the Pro-X-Ser/Thr-Pro MAP kinase substrate consensus sequence phosphorylation site.MAP kinase substrate (MBP)is a peptide MAP kinase substrate (MBP) is a peptide substrate for ERK 1 and ERK 2 MAP kinases. Sequence contains amino acids 95-98 of myelin basic protein (MBP) including Thr 97 within the Pro-X-Ser/Thr-Pro MAP kinase substrate consensus sequence phosphorylation site.MAP kinase substrate (MBP)is a peptide substrate for ERK 1 and ERK 2 MAP kinases... Read More | Inquire | Purity: >90%, by SDS-PAGE visualized with Coomassie® Blue Staining.Description：KGF (keratinocyte growth factor), also known as FGF-7 (fibroblast growth factor-7), is one of 22 known members of the mouse FGF family of secreted proteins that plays a key role in development, Purity: >90%, by SDS-PAGE visualized with Coomassie® Blue Staining.Description：KGF (keratinocyte growth factor), also known as FGF-7 (fibroblast growth factor-7), is one of 22 known members of the mouse FGF family of secreted proteins that plays a key role in development, morphogenesis, angiogenesis, wound healing, and tumorigenesis (1-4). KGF expression is restricted to cells of mesenchymal origin. When secreted, it acts as a paracrine growth factor for nearby epithelial cells (1). KGF speeds wound healing by being dramatically upregulated in response to damage to skin or internal structures that results in high local concentrations of inflammatory mediators such as IL-1 and TNF-alpha. (2, 5). KGF promotes cell migration and invasion, and mediates melanocyte transfer to keratinocytes upon UVB radiation (6, 7). It has been used ectopically to avoid chemotherapy-induced oral mucositis in patients with hematological malignancies (1). Deletion of KGF affects kidney development, producing abnormally small ureteric buds and fewer nephrons (8). It also impedes hair follicle differentiation (9). The 194 amino acid (aa) KGF precursor contains a 31 aa signal sequence and, like all other FGFs, an ~120 aa beta -trefoil scaffold that includes receptor- and heparin-binding sites. KGF signals only through the IIIb splice form of the tyrosine kinase receptor, FGF R2 (FGF R2-IIIb/KGF R) (10). Receptor dimerization requires an octameric or larger heparin or heparin sulfate proteoglycan (11). FGF-10, also called KGF2, shares 51% aa identity and similar function to KGF, but shows more limited expression than KGF and uses an additional receptor, FGF R2-IIIc (12). Following receptor engagement, KGF is typically degraded, while FGF-10 is recycled (12). Mature human KGF, which is active across species, shares 98% aa sequence identity with bovine, equine, ovine and canine, 96% with mouse and porcine, and 92% with rat KGF, respectively... Read More |