Description:
Anti-Mouse IgG2b VHH-Tn5 is the Anti-Mouse IgG2b VHH antibody fused with Tn5. Anti-Mouse IgG2b VHH-Tn5 can bind the mouse IgG2b antibody by specifical VHH antibody, and the fused Tn5 component can provide the function of Tn5 transposons. The specifical VHH has higher affinity than protein A or protein G. the specifical VHH recognizes mouse IgG2b only, and has no cross-reactivity with mouse IgG1/2a/3, rabbit, human, cynomolgus, rat, goat IgG. This product is highly purified to remove contaminating E. coli DNA. This Tn5 is uncharged and must be loaded with user-designed mosaic adapter DNA prior to use in CUT&Tag.
Protein: VHH antibody fused with Tn5
Antibody Specificity: Recognize mouse IgG2b, No cross-reactivity with mouse IgG1/2a/3, rabbit, human, cynomolgus, rat, goat IgG
Purity: Recombinant Expression and Affinity purified
Concentration: 2mg/ml
Molecular weight: 69 kDa
Formation: Liquid, 100 mM Tris-HCl pH 8.0, 0.2 M NaCl, 0.2 mM EDTA, 20% glycerol, 0.2% Triton X-100, 2 mM DTT)
Storage: Store at –20 °C(Avoid freeze / thaw cycles)
Background:
Transposons are genetic elements that can “jump” to different locations within a genome. Bacterial transposons can be divided into the following categories: Insertion sequences, Composite transposons, TnA family, and Muphage. Tn5 is a compound transposon. Tn5 transposons were discovered in Escherichia cdi and consist of a core sequence encoding three antibiotics (neomycin, bleomycin, and streptomycin) and two inverted IS50 sequences, IS50L and IS50R, which encode a Tn5 transposase (Tnp). IS50 has two pairs of 19-bp inverted ends that are outside ends (OEs) and inside ends (IEs). These inverted OEs are target sites of Tn5 transposase. When transposition occurs, transposases bind to the OEs of the Tn5 transposon, forming Tnp-OE complexes, and then the two complexes join together. The C-terminus of Tnp interacts and dimerizes to form a synaptic complex that has the ability to cleave DNA. Tnps that bind to the right and left ends are responsible for catalyzing the hydrolysis of the phosphodiester bond at the left and right ends, respectively. Tnp activates water molecules that hydrolyze the DNA strand and forms a 3′-OH nucleophilic group at the 5′ ends of transposons, which in turn attacks the complementary strand to form a hairpin structure, that further forms a blunt-end by another activated water molecule. Finally, the synaptic complex captures target DNA and finishes the strand transfer by nucleophilic attack on both strands of the target DNA with 3’ OH groups of the Tn5 transposon.
VHH are single-domain antibodies derived from the variable regions of heavy chain of Camelidae immunoglobulin. The size of VHH is extremely small(<15KDa) compared to other forms of antibody fragment, which significantly increase the permeability of VHH. Thus VHH is considered of great value for research, diagnostics and therapeutics.
Fragmentation and adapter addition during the construction of next-generation sequencing (NGS) libraries; Introducing sequencing primers into cloned DNA or plasmids; Establishment of bacterial gene knockout library; Engineering transformation of new bacterial strains; Insertion inactivation of target genes; Insert T7 transcription promoter, resistance markers, etc. into target DNA, etc.
