The binding relation of miR-506-3p to CCL2 was substantiated through RNA immunoprecipitation (RIP) analysis, coupled with the Magna RIP Kit (Millipore). Put simply, NSC-34 and BV2 cells were lysed. Agarose beads (Bio-Rad, Hercules, CA, USA), pre-coated with Argonaute-2 antibody (anti-Ago2), were given for incubating the cell lysates. The antibody of immunoglobulin G (anti-IgG) was employed as the control of our research. Later, the RNA samples combined with the beads were separated and purified. qRT-PCR was carried out to check the abundance of miR-506-3p and CCL2 in immunoprecipitation compounds.
Agarose beads
Manufactured by Bio-Rad
Sourced in United States
Agarose beads are a type of chromatography resin used for the separation and purification of biomolecules such as proteins, nucleic acids, and other macromolecules. They are composed of cross-linked agarose, a polysaccharide derived from certain red algae. Agarose beads have a porous structure that allows for the selective binding and elution of target molecules based on their size, charge, or other physical and chemical properties.
Automatically generated - may contain errors
Lab products found in correlation
Magna rip kit, by Merck Group (2 mentions)
U73122, by Bio-Techne (1 mentions)
Sp600125, by Bio-Techne (1 mentions)
Kn 93, by Bio-Techne (1 mentions)
Brain derived neurotrophic factor (bdnf), by Bio-Rad (1 mentions)
Gf109203x, by Bio-Techne (1 mentions)
Ly294002, by Bio-Techne (1 mentions)
U0126, by Bio-Techne (1 mentions)
Laemmli buffer, by Bio-Rad (1 mentions)
β mercaptoethanol, by Merck Group (1 mentions)
Protein a and g agarose beads, by Thermo Fisher Scientific (1 mentions)
7 protocols using agarose beads
1
miR-506-3p Binding to CCL2 Confirmed
2
Reconstructing Human-Mouse Chimeric Tooth Germs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue recombination and mouse subrenal culture were carried out as described previously [22 (link)]. Briefly, mandibular molar tooth germs dissected from E13.5 mouse embryos were incubated in 2.25% trypsin and 0.75% pancreatin in PBS on ice for 10 min and then dental epithelia were removed with fine forceps. Pieces of confluent hKSC sheets were recombined with E13.5 mouse dental mesenchyme to reconstruct human–mouse chimeric tooth germs [22 (link)]. Agarose beads (Bio-Rad) soaked with FGF8 (125 ng/μl; R&D Systems) and/or SHH (250 ng/μl; R&D Systems), respectively, were implanted into tissue recombinants as described previously [22 (link)]. BSA beads were used as negative control. Recombinant tooth germs were cultured in Trowell type organ culture for 24 h prior to being subjected to subrenal culture in immune-compromised adult male mice. Samples were harvested at various time points after subrenal culture and processed for histological analysis and immunohistochemical staining.
3
Targeted Delivery of Signaling Modulators
4
Immunoprecipitation-based Antigen Discovery
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sera showing moderate or strong reactivity against peripheral nerve cells were used for IP experiments using the same cell as IP substrate. Protein A and G agarose beads (Invitrogen, CA, USA) were used to isolate sera IgG bound overnight to the antigens in a cell culture extract. Precipitated proteins were detached from the agarose beads with Laemmli buffer (Bio-Rad, CA, USA) with 5% b-mercaptoethanol (Merck, Germany) and separated by electrophoresis. Bands appearing in patients’ IP but not in control’s were analyzed by mass spectrometry. Proteins were selected as candidate antigens when they fulfilled any of these criteria: protein score > 100, peptide sequence coverage >5% or two or more peptides identified with the absence of the same criteria in the control sample.
A subset of five patients with typical CIDP that did not react against nerve cells was used for IP experiments using rat whole nerve lysate or human cauda equina as the IP substrate following the exact same protocol to analyze precipitated proteins.
A subset of five patients with typical CIDP that did not react against nerve cells was used for IP experiments using rat whole nerve lysate or human cauda equina as the IP substrate following the exact same protocol to analyze precipitated proteins.
5
Localized SAG Delivery in Rodents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Beads were implanted into the epiphyses of 30-day-old Wistar-Kyoto rats as follows: agarose beads (Bio-Rad, 1537302) incubated with SAG (7 μg in 1.5 μL of DMSO) were placed immediately above the growth plate, in the SOC of the distal femur only. The contralateral hind leg received DMSO-containing beads as a control. The site of administration was accessed from the medial side through a hole drilled with a 22g needle. Following injection, this hole was closed with a hemostatic sponge and the site sutured. In the case of bead implantation into Gli1-LacZ mice, beads incubated with SAG (2.5 mg/mL) or DMSO were injected subcutaneously (amounting ~10 μL bead solution per administration) into the right and left hind paw, respectively, and the animals were sacrificed 1–4 weeks later.
6
Spatio-Temporal Drug Delivery in Zebrafish
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Spatial and temporal control of delivery of pharmacological agents was achieved using agarose beads (100 to 200 mesh, Bio-Rad) loaded with 2 mM Ca2+ medium with or without drugs and implanted at 19 hpf (stage 18) (6 (link), 9 (link), 13 (link)). See SI Appendix for further details.
7
Ago2-Mediated circNINL Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RIP was implemented according to the procedures of Magna RIP™ Kit (Millipore, MA, USA). Cell lysates were separately incubated with Ago2 antibody and IgG antibody and added with agarose beads (Bio-Rad, CA, USA). circNINL, miR-3918 and FGFR1 expression was detected by RT-qPCR [22 (link)].
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!