Protein-heme complexes were prepared by incubating a 120 μM stock solution of human albumin (Sigma; A-8763), Bvu or HmuY in PBS at 37 °C with heme at a 1:1.2 protein-to-heme molar ratio and subsequently passed through Zeba Spin Desalting columns (Thermo Fisher, Scientific, Waltham, MA, USA) to ensure that no uncomplexed heme remained. Coincubation of Bvu with HmuY or with human serum albumin was carried out in PBS at 37 °C and monitored by UV-visible spectroscopy under air (oxidizing) or reducing conditions using each protein at 5 µM concentration [23 (link),24 (link),35 (link)].
A8763
Manufactured by Merck Group
Sourced in United States
A8763 is a laboratory equipment product. It is designed for use in scientific research and testing applications. The core function of this product is to perform specific tasks required for laboratory operations. No further details on the intended use or interpretation of the product's capabilities are provided.
Automatically generated - may contain errors
Lab products found in correlation
Zeba spin desalting column, by Thermo Fisher Scientific (1 mentions)
Copper 2 chloride, by Merck Group (1 mentions)
Chelex 100 resin, by Bio-Rad (1 mentions)
Orion star a211 ph benchtop meter, by Thermo Fisher Scientific (1 mentions)
G9885, by Merck Group (1 mentions)
α1 acid glycoprotein, by Merck Group (1 mentions)
Bixin, by Merck Group (1 mentions)
Vacutainer tube, by BD (1 mentions)
5 protocols using a8763
1
Preparation and Characterization of Protein-Heme Complexes
2
Preparation of Copper-Albumin Solutions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lyophilized fatty acid and globulin-free human serum albumin (rHSA; Sigma-Aldrich A3782) and globulin-free human serum albumin (fHSA; Sigma-Aldrich A8763) were dissolved in 50 mM HEPES pH 7.4 or PBS pH 7.4 buffers. Copper(ii ) chloride (99.9% trace metals basis) was purchased from Sigma (CAS number: 7447-39-4). A stock solution of 0.1 M was prepared using filtered double-distilled H2O treated with Chelex 100 resin (Bio-Rad 1422822).
3
Protein-Ligand Interaction Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bixin (05989, ≥90%, CAS number 6983-79-5), albumin from human serum (A8763, lyophilized powder, essentially globulin free, ≥99%, CAS number 70024-90-7) and α1-acid glycoprotein from human plasma (G9885, ≥99%, CAS number 66455-27-4) were received from Sigma-Aldrich (St. Louis, MO) and utilized without further purification, and deionized water was generated by a Milli-Q Ultrapure Water Purification Systems from Millipore (Billerica, MA). Tris (0.2 M)–HCl (0.1 M) buffer of pH = 7.4, with an ionic strength 0.1 in the presence of NaCl, and the pH was determined with an Orion Star A211 pH Benchtop Meter (Thermo Scientific, Waltham, MA). Dilutions of the protein stock solution (20 μM) in Tris–HCl buffer were prepared immediately before use, and the concentration of proteins was measured by the method of Lowry et al.19 (link) All other reagents used were of analytical grade and purchased from Sigma-Aldrich.
4
Humeral Serum Albumin Challenge in Calves
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Measurements related to performance and health were performed as described above. In addition, one measurement as part of the characterization of the calves from the category immunology was included because this measurement could not be performed earlier for logistic reasons. Calves were subjected to a human serum albumin (HuSa ) challenge in wk 11. Calves with a rectal temperature >40°C were excluded. The HuSa (A8763; Sigma-Aldrich) was dissolved in saline solution at a concentration of 1 mg/mL and was injected intramuscularly at 50 μg/kg of BW. Blood samples were collected in lithium heparin tubes by venipuncture before HuSa injection and at 3, 7, 14, and 28 d after HuSa injection. Plasma was harvested after centrifugation and stored at −20°C pending specific IgG and IgM titer analyses.
5
Calf Antibody Titers after Immunization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood samples of female calves (n = 63) were taken from the jugular vein immediately after birth and every Wednesday in wk 1, 2, 4, 6, 7, 8, 10, 11, and 12 of life in Vacutainer tubes (10 mL; Becton Dickinson and Co., Franklin Lakes, NJ) containing heparin to determine titers of total (IgT), IgG, and IgM natural and specific antibody titers binding KLH or HuSA. Blood samples were centrifuged at 3,000 × g at 4°C for 15 min immediately after sample collection, and plasma was stored at -20°C until analysis. Female calves were immunized twice with a combination of both KLH and HuSA. Solutions of KLH and HuSA were prepared by dissolving 2 mg of KLH (H7017, Sigma Aldrich Co., St Louis, MO) and 0.2 mg of HuSA (A8763, Sigma Aldrich) in 2.5 mL of PBS (0.1 M, pH = 7.2). Calves received a 2.0-mL i.m. injection in the neck at wk 6 of life and a second injection at wk 10 of life.
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!