Blood samples were collected (jugular vein) at day 7 and 14 postpartum and centrifuged (at 3000 rpm for 15 minutes) for serum collection and analysis of BHB and glucose levels, respectively. BHB concentrations in serum were analyzed using the Randox D-3 Hydroxybutyrate (Ranbut) assay and RX monza analyzer for quantitative in vitro determination of D-3-hydroxy butyrate (Ranbut; Randox Laboratories, Antrion, United Kingdom) [23 (link)]. An elevation of BHB concentration above 1.2 mmol/L (12.49 mg/dL) indicates subclinical ketosis in dairy cows [24 (link)]. Glucose was assayed by colorimetric method using a commercial Kit from Biodiagnostic Company (Dokki, Giza, Egypt) and UV-2100 UV/VIS Spectrophotometer (Shimadzu Corporation, Tokyo, Japan) [25 (link)]. Blood samples were classified according to its glucose level as hypo- (<40 mg/dL), normo- (40–60 mg/dL) or hyperglycemic (>60 mg/dL) [26 (link)].
Ranbut
Manufactured by Randox
Sourced in United Kingdom
The Ranbut is a highly specialized laboratory equipment designed for the quantitative measurement of butyrylcholinesterase (BChE) levels in biological samples. It utilizes advanced enzymatic analysis techniques to provide accurate and reliable results.
Automatically generated - may contain errors
Lab products found in correlation
Asys expert plus, by Harvard Bioscience (1 mentions)
Glucose, by Randox (1 mentions)
Hr series nefa hr 2, by Fujifilm (1 mentions)
Osr6134, by Beckman Coulter (1 mentions)
Nefa c, by Fujifilm (1 mentions)
Vacutainer, by BD (1 mentions)
Gluc3, by Roche (1 mentions)
Vitros 5.1 fs, by Ortho Clinical Diagnostics (1 mentions)
Vacutainer vacuum system, by BD (1 mentions)
Cobas bio, by Roche (1 mentions)
Cobas c701 702, by Roche (1 mentions)
Nefa fa115, by Randox (1 mentions)
Ab108842, by Abcam (1 mentions)
Human tnf α quantikine elisa, by R&D Systems (1 mentions)
Human fibrinogen elisa kit, by Abcam (1 mentions)
Serum triglyceride determination kit, by Merck Group (1 mentions)
Vacuette, by Greiner (1 mentions)
Uv 2100 uv vis spectrophotometer, by Shimadzu (1 mentions)
Rx monza analyzer, by Randox (1 mentions)
11 protocols using ranbut
1
Postpartum Serum BHB and Glucose Levels
2
Quantification of Liver Triglycerides in Biological Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum concentrations of glucose (Glucose, Erba mannheim®, UK), BHBA (Ranbut, Randox Laboratories Ltd., UK), NEFA (NEFA, Randox Laboratories Ltd., UK), and IGF-I (human IGF-I, Mediagnost®, Germany) were measured using the commercially available test kits as indicated. Liver concentrations of TAG were measured using the method described previously by van den Top et al. (1994) (link). Briefly, liver tissues were dissolved overnight with 0.5 ml of KOH (20%) and then saponified with 1 ml of absolute ethanol. The reaction tubes were placed in a water bath at 37°C for 1 h. One milliliter of MgSO4 (0.15 M) was added, followed by centrifugation at 1200×g for 10 min. Thereafter, supernatants were removed and 0.5 ml of KOH (0.5 M) in absolute ethanol was added. The sediment and the remaining ethanol were evaporated in a water bath at 100°C. Consequently, 2.0 ml of HCl (2 M) were added, and the tubes were placed in a water bath at 100°C for 2 hours. Titration with NaOH (5 M) was done until pH of 7.0 was attained. TAG concentrations in the supernatant were determined by a commercial kit (Triglyceride, Erba mannheim®, UK) and calculated into liver TAG concentrations, which were expressed as mg of TAG per gram of liver wet weight).
3
Quantification of 3-Hydroxybutyrate in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
3-hydroxybutyrate was measured by a standard method involving conversion of NAD + to NADH determined as the rate of increase of absorbance at 340 nm. A commercial kit RANBUT (Randox Laboratories Limited, Crumlin, UK) was used. Conditioned media (900 µl) were lyophilized and resuspended in 100 µl of ultrapure water. A 50 µl aliquot was used to quantify 3-hydroxybutyrate levels. Cells were disrupted by ultrasonic irradiation in 500 µl of EDTA (2mM) and a 50 µl aliquot was saved for DNA determination. Results were expressed as nmol 3-hydroxybutyrate/µg DNA.
4
Peripartum Plasma BHBA Dynamics in Dairy Cows
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood samples were collected from all cows on days −18 ± 3, −11 ± 3, −4 ± 3, 3 ± 3, 10 ± 3, 17 ± 3, and 24 ± 3 relative to calving from the coccygeal vein or artery immediately after feeding while cows were restrained in self-locking headlocks. Samples were collected into evacuated tubes containing K2 EDTA (Becton Dickinson Vacutainer Systems, Franklin Lakes, NJ, USA). Tubes were placed in ice until centrifugation for plasma separation (1,200 × g for 15 min at 4°C). Plasma was aliquoted into microcentrifuge tubes and stored at −32°C until analysis.
Concentrations of beta-hydroxybutyrate (BHBA) were determined enzymatically [Ranbut, Randox Laboratories, Antrim, UK; (18 (link))] from samples collected weekly from days in milk (DIM) 3 to 24.
Concentrations of beta-hydroxybutyrate (BHBA) were determined enzymatically [Ranbut, Randox Laboratories, Antrim, UK; (18 (link))] from samples collected weekly from days in milk (DIM) 3 to 24.
5
Cow Blood Sampling and Metabolic Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Study personnel collected blood samples from all cows at 3 ± 3, 10 ± 3, and 17 ± 3 DIM using evacuated tubes (Becton Dickinson Vacutainer Systems, Franklin Lakes, NJ) that were placed in ice after collection until centrifugation for serum separation (1,500 × g for 15 min at 4°C). Then, serum was separated and 2 samples were stored in microcentrifuge tubes at -80°C until analyses.
Blood samples collected between 0 and 4 DIM were used to determine total serum calcium concentration using an enzymatic assay (Randox Calcium-CPC/AMP, Randox Laboratories, Antrim, UK). Samples collected between 4 and 20 DIM were used to determine BHB concentrations enzymatically (Ranbut, Randox Laboratories). Cows with Ca concentration ≤8.59 mg/dL were classified as having subclinical hypocalcemia (Martinez et al., 2012) (link) and cows having a BHB concentration >1,200 µmol/L were classified as having subclinical ketosis. At 7 DIM, study personnel evaluated the uterine discharge of cows using the Metricheck device (Simcro Datamars, Hamilton, New Zealand) to determine the percentage of cows with uterine discharge = 5 (fetid, watery, brown/pink; Chenault et al., 2004) (link). At calving and 77 ± 3 DIM, cows were scored for body condition (1 = emaciated and 5 = obese, 0.25-unit increments; Ferguson et al., 1994) (link) by study personnel.
Blood samples collected between 0 and 4 DIM were used to determine total serum calcium concentration using an enzymatic assay (Randox Calcium-CPC/AMP, Randox Laboratories, Antrim, UK). Samples collected between 4 and 20 DIM were used to determine BHB concentrations enzymatically (Ranbut, Randox Laboratories). Cows with Ca concentration ≤8.59 mg/dL were classified as having subclinical hypocalcemia (Martinez et al., 2012) (link) and cows having a BHB concentration >1,200 µmol/L were classified as having subclinical ketosis. At 7 DIM, study personnel evaluated the uterine discharge of cows using the Metricheck device (Simcro Datamars, Hamilton, New Zealand) to determine the percentage of cows with uterine discharge = 5 (fetid, watery, brown/pink; Chenault et al., 2004) (link). At calving and 77 ± 3 DIM, cows were scored for body condition (1 = emaciated and 5 = obese, 0.25-unit increments; Ferguson et al., 1994) (link) by study personnel.
6
Colorimetric Assay for Blood BHBA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BHBA concentrations in the blood serum were determined using the colorimetric method with a reagents kit Ranbut (Randox Laboratories, Crumlin, Antrim, United Kingdom). Intra- and inter-assay coefficients of variation (CV) were 2.8% and 4.7%, respectively. The absorbance readings and subsequent calculations of final concentrations were performed on an automatic microplate reader (Asys Expert Plus, Biochrom Ltd., Cambridge, England) at 450 and 630 nm, respectively.
7
Serum Metabolite Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
8
Serum Biomarkers for Protein and Energy Status
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum samples collected on d 0, 12, and 42 of supplementation were pooled by pasture group and analyzed for BUN, NEFA, and BHBA concentrations to indicate excess protein intake and reduced energy status. Blood was collected using a 38-mm needle into a 10-mL Vacutainer (Becton, Dickinson and Co., Franklin Lakes, NJ) and stored on ice immediately. Blood in the serum tube was allowed to clot at room temperature before being centrifuged (Model HN-S, International Equipment Company, Needham Heights, MA) at 1,300 × g for 20 min at 5°C. About 200 μl from each animal within a group was pooled. Pooled serum was immediately frozen at −20°C until analysis. Composited serum samples were delivered to the University of Illinois, College of Veterinary Medicine Diagnostic Laboratory. Serum BHBA, NEFA, and BUN concentrations were measured in duplicate and analyzed using an Olympus AU680 Chemistry-Immuno Analyzer (Olympus Corporation, Center Valley, PA). Analysis of NEFA was conducted via a colorimetric assay (HR Series NEFA-HR (2); Wako Chemicals, Richmond, VA). Colorimetric analyses were also used for BUN (OSR6134; Beckman Coulter, Indianapolis, IN) and BHBA concentration determination (Ranbut; Randox, Crumlin, UK). The intra- and inter-assay CV were, respectively, 0.8 and 4.9 for NEFA, 2.4 and 2.5 for BUN, and 3.8 and 5.1 for BHBA.
9
Peripartum Metabolic Profiles in Dairy Cows
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood samples were taken 8, 4, 2, and 1 wk prepartum and 1, 2, 3, 4, 6, 9, and 12 wk postpartum. Blood was harvested from the coccygeal artery or vein into heparinized 10-mL vacuum tubes with lithium heparin as anticoagulant (BD Vacutainer, Becton, Dickinson and Company, Franklin Lakes, NJ) at approximately 1000 h each time. The samples were centrifuged at +4°C within 1 h of sampling and plasma samples were stored at -20°C until further analysis. After thawing, plasma samples were analyzed for glucose using enzymatic colorimetric tests (Glucose LiquiColor, Human, Wiesbaden, Germany) and spectrophotometry (Ultrospec K, Boule Nordic, Huddinge, Sweden). The concentration of insulin was analyzed using a commercial enzyme immunoassay method adapted for bovines (Mercodia Ultrasensitive Bovine Insulin ELISA, Mercodia AB, Uppsala, Sweden). The concentration of NEFA was analyzed using an enzymatic colorimetric test (NEFA C, Wako Chemicals GmbH, Neuss, Germany). The concentration of IGF-1 was determined using a commercial enzyme immunoassay (Bovine Insulin-like Growth Factor 1 ELISA catalog no. KT-18278, Kamaya Medical Company, Seattle, WA). The concentration of BHB in plasma was analyzed with a kinetic enzymatic method (RANBUT, Randox Laboratories Limited, Crumlin, UK) and was only analyzed for the 45 cows sampled during yr 2.
10
Fasting Blood Metabolic Profiles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood samples were collected in fasting state from the antecubital vein (BD Vacutainer® vacuum system, Franklin Lakes, NJ, USA), after approximately 8 h of sleep (last meal no more than 2 h before bedtime). The following were determined in the blood: glucose concentration (GLU), ketone bodies (KB) (β-hydroxybutyrate—BHB), triglycerides (TG) and free fatty acids (FFA). GLU concentration was determined in the plasma (EDTA and glycolysis inhibitors: sodium fluoride and potassium oxalate). Triglycerides (TG), FFA and BHB, were determined in the serum (clotting activator). The assays were carried out using commercial reagent kits according to the procedure indicated by the manufacturers.
The detection range was, respectively: 0.11–41.6 mmol/L for glucose (GLUC3 Roche Diagnostics International Ltd., Germany), 0.07–2.24 mmol/L for FFA (NEFA FA 115, Randox Laboratories Ltd., UK), 0.1–5.75 mmol/L for 3-hydroxybutyrate (Ranbut, Randox Laboratories Ltd., UK) and 0.11–5.93 mmol/L for triglycerides (TRIG, Ortho-Clinical Diagnostics, France). The determinations were performed using the Cobas c 701/702 (glucose) and Cobas Bio (FFA, BHB) analysers by Roche Diagnostics International Ltd. (Germany) and Vitros 5.1 FS (TG) by Ortho-Clinical Diagnostics (France). The intra-assay % coefficients of variation were: KB 5.2%, TG 1.1%, FFA 4.3%, GLU 1.1%.
The detection range was, respectively: 0.11–41.6 mmol/L for glucose (GLUC3 Roche Diagnostics International Ltd., Germany), 0.07–2.24 mmol/L for FFA (NEFA FA 115, Randox Laboratories Ltd., UK), 0.1–5.75 mmol/L for 3-hydroxybutyrate (Ranbut, Randox Laboratories Ltd., UK) and 0.11–5.93 mmol/L for triglycerides (TRIG, Ortho-Clinical Diagnostics, France). The determinations were performed using the Cobas c 701/702 (glucose) and Cobas Bio (FFA, BHB) analysers by Roche Diagnostics International Ltd. (Germany) and Vitros 5.1 FS (TG) by Ortho-Clinical Diagnostics (France). The intra-assay % coefficients of variation were: KB 5.2%, TG 1.1%, FFA 4.3%, GLU 1.1%.
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!