NF157, MRS2500, suramin, AR-C 118925XX, CGS15943, pertussis toxin (PTX), ATP, and ATPγS were from Tocris Bioscience (Bristol, UK). YM-254890 (YM) was from Wako Pure Chemical Industries (Richmond, VA). Bovine serum albumin (BSA) fraction V fatty acid-free was from Roche (Basel, Switzerland). Poly-d -lysine and apyrase were from Sigma-Aldrich (St. Louis, MO). Carbachol was from Abcam (Cambridge, UK). Triton X-100 was from Polysciences (Warrington, PA). RANTES was from Peprotech (Rocky Hill, NJ) and RANTES analog, PSC-RANTES was a gift from Oliver Hartley (Université de Genève). HEK293T cells were from American Type Culture Collection (ATCC) (Manassas, VA). Dulbecco’s Modified Eagle’s Medium GlutaMAX (DMEM), FluoroBrite DMEM, Dulbecco’s phosphate-buffered saline without Ca2+ and Mg2+ (DPBS), Hanks' Balanced Salt solution (HBSS), and HEPES buffer were from Fisher Scientific (Hampton, NH). Lipofectamine 2000 and trypsin–EDTA (0.25%, phenol red) were from ThermoFisher Scientific (Waltham, MA). Fetal bovine serum (FBS) was from Gemini Bio-Products (West Sacramento, CA). Clear and clear-bottom black 384-well microplates were from Greiner (Monroe, NC). FLIPR Calcium 6 Assay and Flexstation II 384 Plate Reader were from Molecular Devices (San Jose, CA). 384-well transfer tips for the Flexstation were from Axygen (Union City, CA).
Bovine serum albumin fraction 5 fatty acid free
Manufactured by Roche
Sourced in United States
Bovine Serum Albumin Fraction V, fatty acid free is a laboratory reagent derived from bovine blood. It is a highly purified form of serum albumin protein used in various research and diagnostic applications.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin fraction 5, by Roche (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Poly d lysine, by Merck Group (1 mentions)
Hek293t cell, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Gemini Bio (1 mentions)
Hepes buffer, by Thermo Fisher Scientific (1 mentions)
Rantes, by Thermo Fisher Scientific (1 mentions)
Apyrase, by Merck Group (1 mentions)
Cgs15943, by Bio-Techne (1 mentions)
Ym 254890 ym, by Fujifilm (1 mentions)
Suramin, by Bio-Techne (1 mentions)
Carbachol, by Abcam (1 mentions)
Ar c 118925xx, by Bio-Techne (1 mentions)
Nf157, by Bio-Techne (1 mentions)
Atpγs, by Bio-Techne (1 mentions)
Triton x 100, by Polysciences (1 mentions)
Mrs2500, by Bio-Techne (1 mentions)
3h myo inositol, by PerkinElmer (1 mentions)
Serotonin hydrochloride 5 ht, by Merck Group (1 mentions)
Multiscreen filter plate, by Merck Group (1 mentions)
5 protocols using bovine serum albumin fraction 5 fatty acid free
1
Calcium Signaling Assay in HEK293T Cells
2
Radioligand Binding Assay for 5-HT Receptors
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[ 3 H]LSD (83.6 Ci/mmol), [ 3 H]myo-Inositol (20.3 Ci/mmol) and [ 14 C]arachidonic acid (57.1 mCi/mmol) were purchased from PerkinElmer Life Science (Waltham, Massachusetts). DTT, clozapine, methysergide maleate and serotonin hydrochloride (5-HT) were purchased from Sigma Aldrich (St. Louis, Missouri). RNA Binding YSi SPA Beads and OptiPhase Supermix scintillation cocktail were purchased from Perkin Elmer (Waltham, Massachusetts). Bovine Serum Albumin (BSA) Fraction V fatty acid free was purchased from Roche (Basel, Switzerland). MultiScreen ® Filter Plates were purchased from Millipore (Billerica, Massachusetts). All other reagents were purchased from Sigma Aldrich.
3
Biotinylated Peptide/Protein ELISA
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Example 9
Elisa
To not-coated Maxisorb plates non-biotinylated peptide/protein/aggregate and to streptavidin coated Maxisorb plates biotinylated peptide/protein/aggregate in PBS were added and incubated over-night. The supernatant was discarded and the wells washed three times with 90 μl wash buffer (1×PBS/0.1% Tween 20). Remaining reactive spots were blocked with blocking buffer (1×PBS/2% BSA (Bovine Serum Albumin Fraction V. fatty acid free, Roche, Cat. No.: 10735078001)10.05% Tween 20) by incubating for 1 h. The supernatant was discarded and the wells washed three times with 90 μl wash buffer. Samples and control antibody were prepared in 12 dilutions. (1:2) in ELISA buffer (1×PBS/0.5% BSA (Bovine Serum Albumin Fraction V, fatty acid free, Roche, Cat. No.: 10735078001)/0.05% Tween 20) with a start concentration of 500 ng/mL. The incubation time was 60 minutes at RT on a shaker. The supernatant was discarded and the wells washed three times with 90 μl wash buffer. Solutions of the secondary antibody were prepared in ELISA buffer. A total of 25 μl antibody-mix was transferred in all wells of the assay plate and the plate was thereafter incubated on shaker for 60 minutes at RT. The supernatant was discarded and the wells were washed three times with 90 μl wash buffer. To all wells 25 μl of ABTS solution was added. The absorbance was read at 405 nm-492 nm.
4
Biotinylated Peptide/Protein ELISA
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Example 9
Elisa
To not-coated Maxisorb plates non-biotinylated peptide/protein/aggregate and to streptavidin coated Maxisorb plates biotinylated peptide/protein/aggregate in PBS were added and incubated over-night. The supernatant was discarded and the wells washed three times with 90 μl wash buffer (1×PBS/0.1% Tween 20). Remaining reactive spots were blocked with blocking buffer (1×PBS/2% BSA (Bovine Serum Albumin Fraction V. fatty acid free, Roche, Cat. No.: 10735078001)10.05% Tween 20) by incubating for 1 h. The supernatant was discarded and the wells washed three times with 90 μl wash buffer. Samples and control antibody were prepared in 12 dilutions. (1:2) in ELISA buffer (1×PBS/0.5% BSA (Bovine Serum Albumin Fraction V, fatty acid free, Roche, Cat. No.: 10735078001)/0.05% Tween 20) with a start concentration of 500 ng/mL. The incubation time was 60 minutes at RT on a shaker. The supernatant was discarded and the wells washed three times with 90 μl wash buffer. Solutions of the secondary antibody were prepared in ELISA buffer. A total of 25 μl antibody-mix was transferred in all wells of the assay plate and the plate was thereafter incubated on shaker for 60 minutes at RT. The supernatant was discarded and the wells were washed three times with 90 μl wash buffer. To all wells 25 μl of ABTS solution was added. The absorbance was read at 405 nm-492 nm.
5
Melanocortin Receptor Agonist Protocol
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[Nle 4 ,D-Phe 7 ]-α-melanocyte-stimulating hormone (NDP-MSH) was purchased from Bachem (CA, USA). Sodium palmitate was purchased from Santa Cruz Biotechnology (TX, USA).
Bovine serum albumin, Fraction V, fatty acid free was obtained from Roche (Grenzach, Germany). JKC363 was purchased from Tocris Bioscience (MO, United States). Fetal bovine serum (FBS) was obtained from Natocor (Cordoba, Argentina). Nrf2 antibody (Novus) was kindly provided by Dr. Cymeryng (CEFYBO, UBA-CONICET). DMEM/F-12, DMEM, L-Glutamine and antibiotics were purchased from Invitrogen Life Technologies (CA, USA). All other media and supplements were obtained from Sigma-Aldrich Corporation unless specified otherwise.
Bovine serum albumin, Fraction V, fatty acid free was obtained from Roche (Grenzach, Germany). JKC363 was purchased from Tocris Bioscience (MO, United States). Fetal bovine serum (FBS) was obtained from Natocor (Cordoba, Argentina). Nrf2 antibody (Novus) was kindly provided by Dr. Cymeryng (CEFYBO, UBA-CONICET). DMEM/F-12, DMEM, L-Glutamine and antibiotics were purchased from Invitrogen Life Technologies (CA, USA). All other media and supplements were obtained from Sigma-Aldrich Corporation unless specified otherwise.
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