Oligonucleotide Conjugation Kit (ab218260)

Conjugation - Oligonucleotide Conjugation Kit (ab218260)

Cortez, Valerie, et al used Oligonucleotide Conjugation Kit (ab218260) as part of examining infection of intestine by the murine astrovirus (MuAstV) . They used the kit to conjugate Oligonucleotide to anti-PE antibodies for use in single-cell RNA-sequencing.

Duodenums were collected from infected and uninfected C57BL6 mice at 6 days post MuAstV infection (n?=?4/group) and prepared for single-cell RNA-sequencing. To label cells from each mouse within the infection groups and enable multiplexing, cells were incubated with custom-designed hashtag oligos (HTO) conjugated to anti-PE antibodies using the Oligonucleotide Conjugation Kit (ab218260). The image shows the resulting aggregated data of all duodenal epithelial cells (n?=?2973 individual cells) from both infected and uninfected (n?=?4 mice/group) as represented by t-SNE clustering and colored according to unique cell cluster based on transcriptional profiling.

Image from Cortez et al., Nat Commun., 11(1):2097; doi: 10.1038/s41467-020-15999-y.Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

Oligonucleotide conjugation kit (ab218260) used in studying murine astrovirus (MuAstV) infection in the intestine, by Cortez et al

Conjugation - Oligonucleotide Conjugation Kit (ab218260)

Weigel, Christoph, et al used Oligonucleotide Conjugation Kit (ab218260) as part of examining intragenic methylation loss at the three prime repair exonuclease 2 (TREX2) in laryngeal and colorectal cancer. They used the kit to conjugate DNA oligos to anti-FLAG antibody for use in proximity ligation assay.

Bar chart with relative ligation efficiencies of CEBPA oligonucleotides incubated with protein extract from CEBPA-overexpressing HEK293T cells (CEBPA) versus Empty vector-transfected cell extracts. Mean and standard deviation of quadruplicate experiments is shown. Binding motifs (bold) and respective ligated oligonucleotide sequences are depicted to the left

Image from Weigel et al., Clin Epigenetics, 11(1):67; doi: 10.1186/s13148-019-0666-5. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

Oligonucleotide conjugation kit (ab218260) used in studying methylation loss at TREX2 in laryngeal and colorectal cancer by Weigel et al.

Conjugation - Oligonucleotide Conjugation Kit (ab218260)

A 4-12% reducing gel confirming conjugation between a rabbit IgG and a 30mer oligo done with the Oligonucleotide Conjugation Kit (ab218260). Four different antibody : oligo ratios were tested. 5 μg of conjugate was used for each well.

Reducing gel showing conjugation between rabbit IgG and 30mer oligo using oligonucleotide conjugation kit ab218260

PCR - Oligonucleotide Conjugation Kit (ab218260)

Immuno-qPCR data prepared with the Oligonucleotide Conjugation Kit (ab218260). The graph shows the number of PCR cycles undertaken vs. fluorescence intensity generated by SYBR Green containing PCR probes at particular antigen concentrations. A mouse monoclonal antibody specific for human CRP (clone C7) was purchased in unconjugated format from HyTest. The antibody was conjugated to an oligonucleotide using the Oligonucleotide Conjugation Kit, and was used as a detection antibody in a sandwich immuno-PCR assay with a polyclonal anti-CRP antibody as the capture reagent.

Immuno-qPCR data showing PCR cycles vs. fluorescence intensity using oligonucleotide conjugation kit (ab218260) and SYBR Green probes.

PCR - Oligonucleotide Conjugation Kit (ab218260)

Data from the same experiment was converted to antigen amount vs. cycle number to enable the calculation of a standard curve. The results show that the assay utilizes 1000-fold less capture antibody and 100-fold less detection antibody, and provides 1000-fold greater sensitivity than the equivalent ELISA.