| Description | One unit is defined as the amount of enzyme required to ligate 50% of HindIII fragments of λ DNA (5' DNA termini concentration of 0.12µM, 300µg/ml) in a total reaction volume of 20µl in 30 minutes at 16℃.Application：Ligation of dsDNA with sticky ends; nick repair of One unit is defined as the amount of enzyme required to ligate 50% of HindIII fragments of λ DNA (5' DNA termini concentration of 0.12µM, 300µg/ml) in a total reaction volume of 20µl in 30 minutes at 16℃.Application：Ligation of dsDNA with sticky ends; nick repair of dsDNA; cloning after the synthesis of second strand cDNA [2]. Source：Recombinant E. coli DNA ligase expressed in E. coli.Enzyme storage buffer：10mM Tris-HCl, 50mM KCl, 1mM DTT, 0.1mM EDTA, 200µg/ml BSA, 50% Glycerol (pH7.4, 25℃).Inactivation or inhibition：E. coli DNA Ligase can be inactivated by incubation at 65℃ for 20 minutes.Precautions：The ligation efficiency of this product for blunt-end fragments is extremely low. Use T4 DNA Ligase for blunt-end ligation.This product cannot be used for the ligation of ssDNA or RNA.This kit 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. Set up the reaction on ice as follows:ReagentVolumeFinal ConcentrationdsDNAxµlup to 0.25µg/µl10X Reaction Buffer2µl1XE. coli DNA Ligase (10U/µl)1µl0.5U/µlNuclease-free Water(17-x)µl-Total Volume20µl-Note 1: When multiple reactions are required, prepare a master mix including all reagents except the substrate DNA and then dispense to different nuclease-free PCR tubes. Finally, add substrate DNA to each tube.Note 2: E. coli DNA Ligase should be kept on ice during the experiment.2. Mix well by pipetting or vortex gently. Centrifuge briefly to collect liquid at the bottom of the PCR tube.3. Incubate at 16℃ for 30-60 minutes.4. After incubation, immediately heat inactivate the reaction at 65℃ for 20 minutes.5. Examine the reaction products by agarose gel or polypropylene gel electrophoresis. If DNA need to be recovered from agarose gel, we recommend using the DNA Gel Extraction Kit. To purify DNA from the enzyme digestion reactions, we recommend using the PCR Clean Up Kit/DNA Purification Kit. 6. For other applications, please refer to the relevant literature for E. coli DNA Ligase.FAQ:1. What is the difference between E. coli DNA Ligase and T4 DNA Ligase?Under the recommended conditions in the manual, E. coli DNA Ligase cannot ligate blunt-end DNA or RNA molecules [3].2. Can E. coli DNA Ligase be heat-inactivated?E. coli DNA Ligase can be inactivated by incubation at 65℃ for 20 minutes.3. How to choose the appropriate ligase or ligation kit?For rapid ligation of blunt- or sticky-end DNA fragments, the Rapid DNA Ligation Kit is recommended. T4 DNA Ligase is suitable for most DNA recombination reactions and can be used for sticky-end (10-minute ligation at room temperature) or blunt-end (2-hour or overnight ligation at room temperature) ligations. However, E. coli DNA Ligase is more selective for substrates and is recommended for sticky-end ligations. For ligation of ssDNA or RNA molecules, we recommend using the T4 RNA Ligase. References:1. Lehman IR. Science. 1974. 186(4166):790-7.2. Okayama H and Berg P. Mol Cell Biol. 1982. 2(2):161-70.3. Higgins NP and Cozzarelli NR. Methods Enzymol. 1979. 68:50-71... Read More | Purity:>95%, by SDS-PAGE visualized with Coomassie® Blue StainingDescription:Bcl-2 family proteins contribute to programmed cell death or apoptosis. It is a large protein family and all members contain at least one of four Bcl-2 homology domains. Certain members (Bcl-2, Bcl-XL and Mcl-1) arePurity:>95%, by SDS-PAGE visualized with Coomassie® Blue StainingDescription:Bcl-2 family proteins contribute to programmed cell death or apoptosis. It is a large protein family and all members contain at least one of four Bcl-2 homology domains. Certain members (Bcl-2, Bcl-XL and Mcl-1) are antiapoptotic, whilst others (Bax, Bak, Bok) are proapoptotic... Read More | Purity:>95%, by SDS-PAGE visualized with Coomassie® Blue StainingDescription:Interleukin-6 (IL-6) is a pleiotropic, alpha-helical, 22-28 kDa phosphorylated and variably glycosylated cytokine that plays important roles in the acute phase reaction, inflammation, hematopoiesis, bone metabolism,Purity:>95%, by SDS-PAGE visualized with Coomassie® Blue StainingDescription:Interleukin-6 (IL-6) is a pleiotropic, alpha-helical, 22-28 kDa phosphorylated and variably glycosylated cytokine that plays important roles in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression. Mature human IL-6 is 183 amino acids (aa) in length and shares 39% aa sequence identity with mouse and rat IL-6. Alternative splicing generates several isoforms with internal deletions, some of which exhibit antagonistic properties. IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligand-binding subunit (IL-6 R alpha) and a signal-transducing subunit (gp130). IL-6 binds to IL-6 R alpha, triggering IL-6 R alpha association with gp130 and gp130 dimerization. Gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 R alpha are generated by both alternative splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R alpha elicit responses from gp130-expressing cells that lack cell surface IL-6 R alpha. Trans-signaling enables a wider range of cell types to respond to IL-6, as the expression of gp130 is ubiquitous, while that of IL-6 R alpha is predominantly restricted to hepatocytes, monocytes, and resting lymphocytes. Soluble splice forms of gp130 block trans-signaling from IL-6/IL-6 R alpha but not from other cytokines that use gp130 as a co-receptor. IL-6, along with TNF-alpha and IL-1, drives the acute inflammatory response and the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. When dysregulated, it contributes to chronic inflammation in obesity, insulin resistance, inflammatory bowel disease, arthritis, sepsis, and atherosclerosis. IL-6 can also function as an anti-inflammatory molecule, as in skeletal muscle where it is secreted in response to exercise. In addition, it enhances hematopoietic stem cell proliferation and the differentiation of Th17 cells, memory B cells, and plasma cells... Read More | Purity> 95% by SDS-PAGE and HPLC analyses.FunctionSerine protease inhibitor that inhibits plasminogen activators and plasmin but not thrombin. May be involved in the formation or reorganization of synaptic connections as well as for synaptic plasticity in the adult nervous system. May protect Purity> 95% by SDS-PAGE and HPLC analyses.FunctionSerine protease inhibitor that inhibits plasminogen activators and plasmin but not thrombin. May be involved in the formation or reorganization of synaptic connections as well as for synaptic plasticity in the adult nervous system. May protect neurons from cell damage by tissue-type plasminogen activator... Read More | Purity:>95%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description： 100B, previously called S100 beta, belongs to the S100 family within the EF-hand superfamily of Ca2+ binding proteins. S100 proteins contain two EF-hand motifs that differ in affinity, separated by a hingePurity:>95%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description： 100B, previously called S100 beta, belongs to the S100 family within the EF-hand superfamily of Ca2+ binding proteins. S100 proteins contain two EF-hand motifs that differ in affinity, separated by a hinge region with a hydrophobic cleft that is exposed upon Ca2+ binding. S100B is a 91 amino acid (aa) protein, after removal of the initial methionine, and is found as homodimers of 10.4 kDa monomers. Human S100B shares 99%, 98%, 100%, 99% and 97% aa sequence identity with mouse, rat, rabbit, equine and bovine S100B, respectively. Within the S100 family, human S100B shows the highest aa identity (59%) with S100A1. S100B is expressed primarily by astrocytes and oligodendrocytes in the central nervous system, and by Schwann cells in the peripheral nervous system. Ca2+-bound S100B interacts in vitro with at least 20 cytoplasmic proteins, including several structural molecules such as tubulin and GFAP. It can inhibit the phosphorylation of these kinase substrates and others such as tau and neuromodulin. Astrocytes can secrete S100B, which then acts in a cytokine-like manner. Nanomolar concentrations of S100B are secreted constitutively, promote proliferation, and are neurotrophic and anti-apoptotic. Blood levels of S100B reflect extracellular concentrations within the nervous system, and are elevated in Down’s syndrome, Alzheimer’s disease and Tourette’s syndrome, metabolic stress, acute brain injury and brain tumors. Micromolar concentrations of S100B can be destructive and pro-apoptotic; they induce the expression of iNOS, COX-2, IL-1, IL‑6 and TNF-alpha by microglia, astrocytes or neurons. Most extracellular actions of S100B can be mediated by RAGE (receptor for advanced glycation end products), which is also a receptor for other S100 proteins... Read More |