Biospin tris columns

Manufactured by Bio-Rad

Sourced in United States

The BioSpin Tris Columns are spin columns used for the purification and concentration of biomolecules, such as proteins and nucleic acids, from complex biological samples. These columns are packed with a specialized resin that selectively binds and retains the target biomolecules, allowing for the removal of contaminants and impurities through a simple centrifugation process.

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Lab products found in correlation

Bovine serum albumin (bsa), by Thermo Fisher Scientific (3 mentions) Trichloroacetic acid, by Merck Group (1 mentions) Phosphate buffer saline (pbs), by Thermo Fisher Scientific (1 mentions) Ethyl acetate, by Merck Group (1 mentions) Plga 50 50, by Evonik (1 mentions) C57bl 6 mice, by Jackson ImmunoResearch (1 mentions) Na125i, by PerkinElmer (1 mentions) Trichloroacetic acid tca, by Thermo Fisher Scientific (1 mentions) Fitc conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Non specific igg, by Jackson ImmunoResearch (1 mentions)

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Biospin columns (24 citations)

7 protocols using biospin tris columns

Characterization of Rat Anti-Mouse ICAM-1 Antibody

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Rat monoclonal immunoglobulin G (IgG) Ab against mouse ICAM-1
(anti-ICAM) was clone YN1, produced in a hybridoma from American Type Culture
Collection (Manassas, VA). Non-specific IgG Ab (called IgG hereafter) was from
Jackson Immunoresearch (Pike West Grove, PA). Polyclonal anti-PECAM-1,
FITC-labeled secondary Abs and Alexa-fluor 488 labeled secondary antibodies were
from Novus Biologicals (Centennial, CO), Jackson Immunoresearch (West Grove, PA)
and Thermo fisher Scientific (Waltham, MA), respectively. DNA oligonucleotide
(72-mer) modified with thiol at 5’ was from Oligo Factory (Holliston,
MA). Pierce bond-breaker TCEP solution, LC-SMCC crosslinker, 7k MWCO Zeba spin
columns, 10k MWCO Amicon spin filters, Sartorius™ Vivaspin™ 500
centrifugal concentrator filters (1,000 KDa), thiophilic adsorption resin,
heterobifunctional Pierce crosslinking kit, and bovine serum albumin (BSA) were
from Fisher Scientific (Kerrville, TX). Carboxylic acid-terminated PLGA (50:50
copolymer ratio; 32 kDa average molecular weight) was from Sigma-Aldrich (Saint
Louis, Missouri). Iodogen iodination tubes were from Pierce (Rockford, Illinois)
and BioSpin Tris Columns from BioRad (Hercules, California). All other reagents
were from Sigma Chemical (St. Louis, MO).

Roki N., Tsinas Z., Solomon M., Bowers J., Getts R.C, & Muro S. (2019). Unprecedently High Targeting Specificity Toward Lung ICAM-1 Using 3DNA Nanocarriers. Journal of controlled release : official journal of the Controlled Release Society, 305, 41-49.

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PLGA-based Targeted Drug Delivery

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PLGA 50:50 (ester terminated, 55–80 kDa average molecular weight (MW)) was purchased from Evonik (Birmingham, AL, USA). PLGA 60:40 (ester terminated, 45–55 kDa average MW) and PLGA 75:25 (ester terminated, 45–55 kDa average MW) were purchased from PolySciTech (West Lafayette, IN, USA). Mouse anti-human ICAM-1 (clone R6.5) was affinity purified from a hybridoma obtained from American Type Culture Collection (Manassas, VA, USA). Bovine serum albumin (BSA) and phosphate buffer saline (PBS) were from Thermo Fisher Scientific (Waltham, MA, USA). Pierce Iodogen and ¹²⁵Iodine (¹²⁵I) were from Thermo Fisher Scientific (Waltham, MA, USA). BioSpin Tris Columns were from BioRad (Hercules, CA, USA). HAse Type I-S from bovine testes, 1000–2200 kDa hyaluronic acid (HA), Pluronic F68, ethyl acetate, trichloroacetic acid (TCA), and all other reagents were from Sigma-Aldrich (St. Louis, MO, USA). Gel permeation chromatography (GPC)-grade tetrahydrofuran (THF) was from Scharlab S.L. (Barcelona, Spain).

del Moral M., Loeck M., Muntimadugu E., Vives G., Pham V., Pfeifer P., Battaglia G, & Muro S. (2023). Role of the Lactide:Glycolide Ratio in PLGA Nanoparticle Stability and Release under Lysosomal Conditions for Enzyme Replacement Therapy of Lysosomal Storage Disorders. Journal of Functional Biomaterials, 14(9), 440.

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Immunoglobulin G Labeling and Characterization

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Nonspecific rat immunoglobulin G Ab was from Jackson Immunoresearch (Pike West Grove, PA) and mouse anti‐human ICAM‐1 clone YN.1 was from American Type Culture Collection (Manassas, VA). 72‐mer DNA oligo with 5' thiol modification was from Oligo Factory (Holliston, MA). Pierce Bond‐Breaker TCEP Solution, LC‐SMCC Crosslinker, Zeba Spin Columns (7k MWCO), Thiophilic Adsorption Resin, Amicon 10k MWCO spin filters, and Heterobifunctional Crosslinking Kit were from Fisher Scientific (Kerrville, TX). Iodogen tubes were from Pierce (Rockford, IL) and BioSpin Tris Columns from BioRad (Hercules, CA). Na¹²⁵I was from Perkin‐Elmer (Waltham, MA). All other reagents were from Sigma Chemical (St. Louis, MO). Eight‐week old C57BL/6 mice were from Jackson Laboratory (Bar Harbor, ME). All animal experiments were performed under University of Maryland IACUC approval and adhered to the Principles of Laboratory Animal Care.

Roki N., Solomon M., Casta L., Bowers J., Getts R.C, & Muro S. (2021). A method to improve quantitative radiotracing‐based analysis of the in vivo biodistribution of drug carriers. Bioengineering & Translational Medicine, 6(2), e10208.

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Reoxidation Analysis of RNase A by PDI9, ERO1, and SNO-ERO1

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To denature RNase A, 2 mg RNase A was added into 1 ml 8 M urea containing 150 mm DTT. After 2 h shaking, urea and DTT were replaced by 100 Tris-HAc, pH 8.0, 50 mM NaCl, and 1 mM EDTA. For GSNO incubation, 100 μg recombinant wild type or Cys337 mutated ERO1 proteins were incubated with 2.5 mM GSNO in HEN solution overnight at 4 °C in the dark. Excess GSNO was removed using Bio-Spin Tris columns (Bio-Rad).
Gel-based RNase A reoxidation analysis was performed as described previously^{45 (link)}. 2 μM PDI9, 2 μM ERO1 or SNO-ERO1, 100 μM FAD and 8 μM denatured and reduced RNase A were mixed and incubated in 50 μl reaction buffer (100 Tris-HAc, pH 8.0, 50 mM NaCl, and 1 mM EDTA) at 25 °C. At the indicated time points, the reaction was terminated by the addition of 5 × SDS loading buffer and 10 mM 4-acetamido-4’-maleimidylstilbene-2,2’ disulfonic acid (AMS) to block the free thiols. The samples were analyzed by non-reducing SDS PAGE and stained with Fast Protein Stain (Biofuraw, UK).

Qin G., Qu M., Jia B., Wang W., Luo Z., Song C.P., Tao W.A, & Wang P. (2023). FAT-switch-based quantitative S-nitrosoproteomics reveals a key role of GSNOR1 in regulating ER functions. Nature Communications, 14, 3268.

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Antibody and Reagent Preparation

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Rat anti-mouse ICAM-1 (anti-ICAM) antibody YN1, was produced from respective hybridoma from the American Type Culture Collection (Manassas, VA, USA). Non-specific IgG was from Jackson Immunoresearch (Pike West Grove, PA, USA). 5′-modified DNA oligonucleotide (72-mer) was from Oligo Factory (Holliston, MA, USA). Pierce bond-breaker TCEP solution, LC-SMCC crosslinker, 7 k MWCO Zeba spin columns, thiophilic adsorption resin, heterobifunctional Pierce crosslinking kit, bovine serum albumin (BSA), and trichloroacetic acid (TCA) were from Fisher Scientific (Kerrville, TX, USA). Rabbit anti-mouse PECAM-1 antibody was from Novus Biologicals (Centennial, CO, USA) and FITC-conjugated goat anti-rabbit IgG was from Invitrogen (Carlsbad, CA, USA). Iodogen iodination tubes were from Pierce (Rockford, Illinois) and BioSpin Tris Columns were from BioRad (Hercules, California). Amicon 10 kDa MWCO spin filters were from Millipore Sigma. All other reagents were from Sigma Chemical (St. Louis, MO, USA).

Roki N., Solomon M., Bowers J., Getts L., Getts R.C, & Muro S. (2022). Tuning Design Parameters of ICAM-1-Targeted 3DNA Nanocarriers to Optimize Pulmonary Targeting Depending on Drug Type. Pharmaceutics, 14(7), 1496.

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Radiolabeling and Fluorescent Labeling of Molecules

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Where indicated, anti-ICAM (R6.5 or YN1) or enzymes (HAse or ASM) were labeled with ¹²⁵Iodine (¹²⁵I) or AlexaFluor 488 for radioisotopic quantification or fluorescence tracing, respectively. ¹²⁵I was conjugated to antibody or enzymes by incubation at 1 mg/mL for 5 min at 4 °C with 20 µCi ¹²⁵I using Pierce Iodogen. Free ¹²⁵I was then removed using BioSpin Tris Columns (BioRad; Hercules, CA, USA). Trichloroacetic acid precipitation of the protein followed by (i) centrifugation to separate protein in the pellet vs. free ¹²⁵I in the supernatant and (ii) determination of the label and protein concentration in the pellet, were used to determine the protein specific activity (cpms/mass).
HAse labeling with green, fluorescent AlexaFluor488 was completed by conjugation. Briefly, 1 M sodium bicarbonate was added to 0.5 mL of 2 mg/mL HAse solution to raise the pH up to 8.3. Reactive dye from the labeling kit was then added to the solution and the reaction was stirred at room temperature for 1 h, after which non-conjugated dye was removed by size exclusion chromatography following vendor’s instructions.

Muntimadugu E., Silva-Abreu M., Vives G., Loeck M., Pham V., del Moral M., Solomon M, & Muro S. (2022). Comparison between Nanoparticle Encapsulation and Surface Loading for Lysosomal Enzyme Replacement Therapy. International Journal of Molecular Sciences, 23(7), 4034.

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Radioactive and Fluorescent Labeling of Antibody Conjugates

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Where indicated, anti-ICAM-oligo or IgG-oligo were labeled with ¹²⁵I or Cy3 for radioisotopic quantification or fluorescence tracing, respectively. Conjugation to ¹²⁵I was achieved by incubating 1 mg/mL Ab-oligo for 5 min at 4 °C with 20 µCi ¹²⁵I, using Pierce Iodogen, after which non-conjugated ¹²⁵I was removed using BioSpin Tris Columns (BioRad; Hercules, CA, USA). The specific activity (cpms/mass) of the resulting ¹²⁵I-Ab-oligo was obtained by further separating free ¹²⁵I by precipitation in 15% (v/v) trichloroacetic acid (TCA), followed by centrifugation to separate ¹²⁵I-Ab-oligo in the pellet and quantification of respective cpms and protein concentration, as published [7 (link)].
Fluorescent labeling of 3DNA was pursued using commercial Cy3-oligo conjugates from Integrated DNA Technologies (Coralville, IA, USA), where the fluorophore was located at the 5′-end of an oligo whose sequence was complementary to that of 3DNA arms. The conjugate was hybridized to 3DNA, followed by psoralen crosslinking and purification using size exclusion chromatography. Ab-oligo was then annealed to Cy3-3DNA as described above in Section 2.2.

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