Rnase free bovine serum albumin

Manufactured by Merck Group

Sourced in United States

RNase-free bovine serum albumin is a purified protein used in laboratory applications. It serves as a stabilizer and blocking agent to prevent RNase contamination in RNA-based experiments.

Automatically generated - may contain errors

Lab products found in correlation

Yeast trna, by Merck Group (10 mentions) M 280 streptavidin magnetic bead, by Thermo Fisher Scientific (3 mentions) Streptavidin magnetic beads, by Thermo Fisher Scientific (2 mentions) Trizol, by Merck Group (2 mentions) Biotinylated nc, by GenePharma (2 mentions) Bio mir nc, by GenePharma (1 mentions) Luciferase reporter gene assay kit, by Beyotime (1 mentions) 4 2 hydroxyethyl 1 piperazineethanesulfonic acid hepes, by MP Biomedicals (1 mentions) Rnase free water, by Thermo Fisher Scientific (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions) Perchloric acid, by Merck Group (1 mentions) Lysis buffer, by Thermo Fisher Scientific (1 mentions) Mir 98 5p mut, by Genecreate (1 mentions)

Close spelling variants of this product

Bovine albumin (95 citations) Bovine serum (80 citations) -free bovine serum albumin (8 citations) RNases (8 citations) Serum bovine albumin (7 citations) RNase-free bovine serum albumin and yeast tRNA (6 citations) Albumins ( (2 citations)

10 protocols using rnase free bovine serum albumin

Affinity Purification of Biotinylated miRNA

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HEK293 cells were transfected with biotinylated miRNA (200 nM) and harvested 24 h
after transfection. Briefly, the cells were washed with PBS followed by brief
vortex, and incubated in a lysis buffer on ice for 10 min. The lysates were
precleared by centrifugation, and 50 µl of the samples were aliquoted for input.
The remaining lysates were incubated with streptavidin magnetic beads (Thermo
Fisher Scientific). After coated with RNase-free bovine serum albumin and yeast
tRNA (both from Sigma), the beads were incubated at 4°C for 3 h, washed twice
with ice-cold lysis buffer, three times with the low-salt buffer, and once with
the high-salt buffer. The bound RNAs were purified using TRIzol and then for
NEAT1 expression analysis by RT-qPCR.

Pan W., Wu A., Yu H., Yu Q., Zheng B., Yang W., Tian D., Gao Y, & Li P. (2020). NEAT1 Negatively Regulates Cell Proliferation and Migration of Neuroblastoma Cells by miR-183-5p/FOXP1 Via the ERK/AKT Pathway. Cell Transplantation, 29, 0963689720943608.

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Circular RNA-miRNA Interaction Validation

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Circular RNA interactome (https://circinteractome.nia.nih.gov/mirna_target_sites.html) and circBank (http://www.circbank.cn/) were used to predict the binding sites of miR‐637 for circ_0081054. MicroT CDS (https://dianalab.e‐ce.uth.gr/html/dianauniverse/index.php?r = microT_CDS) was used for predicting the target sites between miR‐637 and RAB9A.RNA pull‐down assay was used to validate the interaction between circ_0081054 and miR‐637. A375 and A2058 cells were transfected with biotinylated circ_0081054 (circ_0081054 probe; GenePharma, Shanghai, China) or biotinylated con (control probe; GenePharma). When the cells were disrupted after 48 h later, the lysate was mixed with magnetic beads (M‐280; Invitrogen) coated with RNase‐free bovine serum albumin (Sigma‐Aldrich) and yeast tRNA (Sigma‐Aldrich) at 4°C for 3 h. Subsequently, bound RNA was purified then detected by qRT‐PCR assay.

Li X., Kong Y., Li H., Xu M., Jiang M., Sun W, & Xu S. (2023). Circ_0081054 facilitates melanoma development via sponging miR‐637 and regulating RAB9A. Skin Research and Technology, 29(5), e13313.

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Biotinylated miRNA Pull-Down Assay

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RNA pull‐down assays were performed as described previously 14. NOZ cells were transfected with biotinylated miR‐363‐3p, biotinylated miR‐363‐3p‐mut and biotinylated NC. Cells were collected at 48 hrs. The cells lysates were incubated with M‐280 streptavidin magnetic beads (Invitrogen, San Diego, CA, USA). To prevent non‐specific binding of RNA and protein complexes, the beads were coated with RNase‐free bovine serum albumin and yeast tRNA (both from Sigma‐Aldrich). The beads were incubated at 4°C for 3 hrs and washed three times with ice‐cold lysis buffer and once with high salt buffer (0.1% SDS, 1% Triton X‐100, 2 mM EDTA, 20 mM Tris‐HCl, pH 8.0 and 500 mM NaCl). The bound RNAs were purified using TRIzol for the analysis.

Wang S., Zhang W., Wu X., Weng M., Zhang M., Cai Q., Zhou D., Wang J, & Quan Z. (2016). The lncRNA MALAT1 functions as a competing endogenous RNA to regulate MCL‐1 expression by sponging miR‐363‐3p in gallbladder cancer. Journal of Cellular and Molecular Medicine, 20(12), 2299-2308.

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Investigating Circular RNA-miRNA Interactions

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Wild‐ and mutant‐type luciferase reporters of hsa_circ_0018289 (hsa_circ_0018289‐WT and hsa_circ_0018289‐MUT) and ICMT (ICMT‐3’UTR‐WT and ICMT‐3’UTR‐MUT) containing the predicted binding sites with miR‐1294 were designed with Hanbio Biotechnology. HeLa and SiHa cells were co‐introduced with a luciferase reporter and miR‐NC or miR‐1294. Luciferase Reporter Gene Assay Kit (Beyotime) was used for measuring luciferase density according to the producer's guidance, with Renilla luciferase activity as control.
For RNA pull‐down assay, CC cells were transfected with biotinylated (Bio)‐miR‐1294 (GenePharma) or Bio‐miR‐NC (GenePharma). 48 h later, cells were disintegrated, and lysate was mixed with magnetic beads (M‐280; Invitrogen) coated with RNase‐free bovine serum albumin (Sigma‐Aldrich) and yeast tRNA (Sigma‐Aldrich) at 4°C for 3 h. After being purified, bound RNA was subjected to the analysis of qRT‐PCR.

Li Y., Gao X., Yang C., Yan H, & Li C. (2022). CircRNA hsa_circ_0018289 exerts an oncogenic role in cervical cancer progression through miR‐1294/ICMT axis. Journal of Clinical Laboratory Analysis, 36(5), e24348.

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Identification of miRNA Targets by Biotin Pulldown

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HEK-293T cells were transfected with biotinylated miRNA (200 nM) for 24 h. The cells were gently washed twice with PBS and lysed by using ice-cold RNA pull-down lysis buffer on ice for 10 min. The lysate obtained in the previous step was centrifuged, and the supernatant was aliquoted into 50 µl for subsequent input research. Streptavidin magnetic beads (Thermo Scientific Fisher Scientific) were added to the remaining lysate and incubated at room temperature. Then RNase-free bovine serum albumin (Sigma) and yeast tRNA (Sigma) were added. The mixture was incubated at 4 °C for 3 h. In order to obtain a pure sample, the beads were washed with ice-cold lysis buffer and low-salt buffer for three times and finally washed with high-salt buffer. At last, TRIzol (Sigma) was used to purify the bound RNAs, and RT-qPCR was used to analyze the expression of HCG11.

Li D., Liu Y., Gao W., Han J., Yuan R., Zhang M, & Ge Z. (2020). LncRNA HCG11 Inhibits Adipocyte Differentiation in Human Adipose-Derived Mesenchymal Stem Cells by Sponging miR-204-5p to Upregulate SIRT1. Cell Transplantation, 29, 0963689720968090.

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Biotinylated miRNA Pulldown Assay

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NOZ cells were transfected with biotinylated miR-138, biotinylated miR-138-mut and biotinylated NC (GenePharma, China). Cells were collected at 48 h. The cell lysates were incubated with M-280 streptavidin magnetic beads (Invitrogen, USA). To prevent non-specific binding of RNA and protein complexes, the beads were coated with RNase-free bovine serum albumin and yeast tRNA (both from Sigma-Aldrich). The beads were incubated at 4°C for 3 h, and washed three times with ice-cold lysis buffer and once with high salt buffer (0.1% SDS, 1% Triton X-100, 2 mM EDTA, 20 mM Tris–HCl, pH 8.0 and 500 mM NaCl). The bound RNAs were purified using TRIzol for the analysis.

Cai Q., Wang Z., Wang S., Weng M., Zhou D., Li C., Wang J., Chen E, & Quan Z. (2017). Long non-coding RNA LINC00152 promotes gallbladder cancer metastasis and epithelial–mesenchymal transition by regulating HIF-1α via miR-138. Open Biology, 7(1), 160247.

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Assessing Angiogenin-GST Ribonucleolytic Activity

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The ribonucleolytic activity of purified wild-type and mutated proteins were examined using yeast transfer RNA (tRNA) as substrate. 0.05–0.5 mg/ml of wild-type and mutant Angiogenin-GST proteins were added to a 300 µl assay mixture containing 0.6 mg yeast tRNA (Sigma), 30 µg RNase-free bovine serum albumin (Sigma), 30 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), pH 6.8 (MP Biomedicals), and 30 mM sodium chloride (Sigma), and the assay mixtures were incubated for 2 hours at 37°C. Subsequently, 700 µl of 3.4% pre-chilled, ice-cold perchloric acid (Sigma) was added to assay mixtures to terminate the reactions. The mixtures were then rigorously vortexed, incubated on ice for 15 minutes, followed by centrifugation at 14000 rpm for 10 minutes at 4°C. The supernatants were collected, and their absorbance measured at 260 nm. The percentage loss of ribonucleolytic activity was calculated by considering the amount of mutated proteins required to generate 1.0 optical density (OD), keeping wild-type Angiogenin-GST as reference. RNAse-free water (Gibco) was used for preparing the buffers and assay mixtures to ensure RNAse-free systems. Experiments were done in triplicates.

Padhi A.K., Banerjee K., Gomes J, & Banerjee M. (2014). Computational and Functional Characterization of Angiogenin Mutations, and Correlation with Amyotrophic Lateral Sclerosis. PLoS ONE, 9(11), e111963.

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Biotinylated miRNA Pulldown in HEK-293T Cells

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HEK‐293T cells were treated for 24 h with biotinylated miRNA (200 nM). Next, the cells were gently rinsed two times with PBS and then lysed for 10 min with ice‐cold RNA pull‐down lysis buffer on ice. The obtained lysate was centrifuged, and the supernatant that was aliquoted into 50 µL was used for subsequent input research. Afterward, streptavidin magnetic beads (Thermo Scientific Fisher) were supplemented to the remaining lysate, followed by incubation at room temperature and addition of RNase‐free bovine serum albumin (Sigma) and yeast tRNA (Sigma). Subsequently, the mixture was incubated for 3 h at 4°C. The beads were rinsed with ice‐cold lysis buffer and low‐salt buffer for three times and lastly rinsed with high‐salt buffer to obtain a pure sample. At last, TRIzol (Sigma) was adopted for purifying the bound RNAs, and RT‐qPCR was carried out to analyze XIST expression.

Bai Q., Li Y., Song K., Huang J, & Qin L. (2021). Knockdown of XIST up‐regulates 263294miR‐340‐5p to relieve myocardial ischaemia–reperfusion injury via inhibiting cyclin D1. ESC Heart Failure, 9(2), 1050-1060.

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Biotinylated miR-203 Pulldown Assay

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Biotinylated miR‐203, biotinylated miR‐203‐mut and biotinylated NC (GenePharma, China) were transfected into H1299 cells and harvested 48 hours later. Cell lysates were incubated with M‐280 streptavidin magnetic beads (Invitrogen, USA) coated with RNase‐free bovine serum albumin (Sigma‐Aldrich) and yeast tRNA (Sigma‐Aldrich) to prevent nonspecific binding of RNA and protein complexes. The beads were incubated at 4 °C for 3 hours. Next, beads were washed three times in ice cold lysis buffer and once in high salt buffer. Bound RNAs were isolated by using TRIzol for subsequent analysis.

Xue Y., Ding M., Xue L, & Luo J. (2019). CircAGFG1 sponges miR‐203 to promote EMT and metastasis of non‐small‐cell lung cancer by upregulating ZNF281 expression. Thoracic Cancer, 10(8), 1692-1701.

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Biotin-labeled miR-98-5p RNA Enrichment

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Biotin-labeled miR-98-5p-WT and miR-98-5p-Mut were synthesized by GeneCreate (Wuhan, China) and were transfected into GC cells which were incubated with lysis buffer (Ambion, Austin, Texas, USA). Then the GC cell lysates were incubated with the streptavidin Dynabeads (Invitgen, USA) precoated with RNasefree bovine serum albumin (BSA) and yeast tRNA (Sigma-Aldrich, USA) overnight at 4 °C. After washed with washing buffer, the bound RNA was puri ed by using Trizol. Finally, the enrichment of PITPNA-AS1 was identi ed and estimated by performing qRT-PCR.

Liu Y., Zhao X., Chen Y., Guo G., Wang J., & He S. (2021). PITPNA-AS1 Activated by H3K27ac Sponged miR-98-5p to Regulate Cisplatin Resistance in Gastric Cancer.

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