MCF-7 cell lysates were incubated with CBR3-AS1 or control probe (Sangon, China) conjugated to streptavidin agarose resin beads (Thermo Fisher Scientific, USA) to generate probe-bound Dynabeads. After the samples were washed with buffer, DNase I (20 U) was added, and purified RNA was collected. The purified RNA was analysed by qRT-PCR. The whole experimental process and the buffer formulation were described by Hsu et al. [24 (link)].
Control probe
Manufactured by Sangon
Sourced in China
The control probe is a laboratory instrument used to maintain and monitor the stability and reliability of analytical measurements. It serves as a reference point for calibrating and validating the performance of other equipment, ensuring accurate and consistent results.
Automatically generated - may contain errors
Lab products found in correlation
Streptavidin coated beads, by Thermo Fisher Scientific (3 mentions)
Trizol, by Takara Bio (2 mentions)
Streptavidin agarose resin beads, by Thermo Fisher Scientific (1 mentions)
Streptavidin coated magnetic beads, by Merck Group (1 mentions)
Unisart cn140, by Sartorius (1 mentions)
Milli q reagent water, by Merck Group (1 mentions)
Streptavidin c1 magnetic beads, by Thermo Fisher Scientific (1 mentions)
7 protocols using control probe
1
Quantifying CBR3-AS1 RNA Interactions
2
Biotin-labeled miR-127-5p Pulldown Assay
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Biotin-labeled miR-127-5p probe and control probe were synthesized by Sangon Biotech (Shanghai, China). Probe-coated beads were generated by co-incubating the probe with streptavidin-coated beads (Invitrogen, CA, USA) at a temperature of 25°C for 2 hours. ESCC cells were collected and lysed and incubated with miR-127-5p probes overnight at 4°C. Following this, the beads were eluted and the complex was purified with TRIzol (Takara, Dalian, China). Then the abundance of circNELL2 and CDC6 was analyzed using qRT-PCR.
3
Biotin-coupled RNA Complex Pulldown
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The biotin‐coupled RNA complex was pulled down by incubating the melanoma cell lysates with streptavidin‐coated magnetic beads (Sigma) following the manufacturer's instructions. qRT‐PCR analysis was used to evaluate the enrichment of BASP1‐AS1 in the captured fractions. Both the BASP1‐AS1 junction probe and the control probe were obtained from Sangon Biotech. The proteins in the capture complex were identified by western blotting using an anti‐YBX1 antibody.
4
Affinity Capture of AK136714 Interactors
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The biotinylated probe of AK136714 and the control probe were synthesized by Sangon Biotech (Shanghai, China). Probe-coated beads were generated by co-incubating the probe with streptavidin-coated beads (Invitrogen, CA, USA) at 25° C for 2 h. Cells were lysed to extract total protein. After pretreatment of magnetic beads, RNA and beads were mixed. After separation, western blot and mass spectrometry were used to verify the downstream binding protein of AK136714.
5
Profiling miR-124-5p and AK003290 Interactions
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Biotinylated miR-124-5p probe and the control probe were synthesized by Sangon Biotech (Shanghai, China). Probe-coated beads were generated by co-incubating the probe with streptavidin-coated beads (Invitrogen, CA, USA) at 25 °C for 2 h. H9c2 cells were collected, lysed, and incubated with AK003290 or miR-124-5p probes overnight at 4 °C. Thereafter, the beads were eluted, and the complex was purified with TRIzol (Takara, Dalian, China). Then, the abundance of AK003290 and miR-124-5p was analyzed by qRT-PCR.
6
Lateral Flow Assay for HIV-1 Detection
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Ball pen (M&G, ball size 1 mm) was purchased from local store. Chloroauric acid (HAuCl4·4H2O) and sodium citrate (Na3C6H5O7·2H2O) were purchased from Shanghai Sinopharm Chemical Reagent Co., Ltd. (China) and American AMRESCO, respectively. The nitrocellulose (NC) membranes (UniSart CN 140) were obtained from Sartorius Stedim Biotech. The conjugate pads, backing pads and absorbent pads used to prepare LFAs were purchased from Shanghai Jiening Biotechnology Co., Ltd. (China). The reagents including detection probe, capture probe, control probe and target DNA used for detection of human immunodeficiency virus type 1 (HIV-1), were synthesized by Shanghai Sangon Biotech Co., Ltd. (China). The detailed sequences were shown in Table 1 . All aqueous solutions used in this work were prepared from Milli-Q reagent water (Millipore Corp., resistivity of 18.2 MΩ cm at 25 °C).
7
circVAMP3 Protein Interaction Assay
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A biotin-labeled circVAMP3 probe targeting the unique circVAMP3 back splicing junction (BSJ) and control probe were purchased from Sangon Biotech (Shanghai, China). They were incubated with streptavidin C1 magnetic beads (Invitrogen, USA) for 15 min at room temperature and then incubated with cell lysates or in vitro transcribed circVAMP3 and purified NP or NS1 protein at 4°C for 4 h. The bound proteins were then detected by immunoblotting.
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