[2,3-3H]D-aspartate (specific activity 11.3 Ci/mmol) was from Perkin Elmer (Boston, MA, USA). (S)AMPA, pep2-SVKI, pep2-SVKE, 2,3-Dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulfonamide disodium salt (NBQX), and DL-threo-β-Benzyloxyaspartic acid (DL-t-BOA) were purchased by Tocris Bioscience (Bristol, UK). Horseradish peroxidase-conjugated anti-mouse and anti-rabbit secondary antibodies and glycine were from Sigma (Milan, Italy). Luminata Forte Western blotting detection system, guinea pig anti-vesicular glutamate transporters type 1 (VGLUT1) antibody and mouse anti-GluA3 monoclonal antibody (MAB5416) were purchased from Millipore (Temecula, CA, USA). Rabbit anti-GluA1 polyclonal antibody (ab86141) and rabbit anti-GluA2 monoclonal antibody (ab206293, EPR18115) were from Abcam (Cambridge, UK). Rabbit anti-GluA4 monoclonal antibody was from GeneTex (GTX62957, Irvine, USA). Donkey anti-rabbit AlexaFluor-647, goat anti guinea pig AlexaFluor-688, donkey anti-mouse AlexaFluor-647, and donkey anti-goat AlexaFluor-688 were from Life Technologies Corporation (Carlsbad, CA, USA). Anti-syntaxin 1A monoclonal mouse IgG, was purchased from Chemicon (CA;USA). Complement and C1q-depleted complement was from Gentaur (Kampenhout, Belgium).
Horseradish peroxidase conjugated anti mouse and anti rabbit secondary antibodies
Manufactured by Merck Group
Sourced in Italy
Horseradish peroxidase-conjugated anti-mouse and anti-rabbit secondary antibodies are laboratory reagents used in various immunoassays and detection techniques. They are designed to bind to primary antibodies raised in mouse or rabbit and enable signal amplification through the enzymatic activity of horseradish peroxidase.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Ab86141, by Abcam (1 mentions)
Ab206293, by Abcam (1 mentions)
2 3 3h d aspartate, by PerkinElmer (1 mentions)
S ampa, by Bio-Techne (1 mentions)
2 3 dioxo 6 nitro 1 2 3 4 tetrahydrobenzo f quinoxaline 7 sulfonamide disodium salt nbqx, by Bio-Techne (1 mentions)
Dl threo β benzyloxyaspartic acid dl t boa, by Bio-Techne (1 mentions)
Alexa fluor 647 donkey anti rabbit, by Thermo Fisher Scientific (1 mentions)
Donkey antimouse alexa fluor 647, by Thermo Fisher Scientific (1 mentions)
Anti caga, by Santa Cruz Biotechnology (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Rabbit anti gapdh antibody, by Abcam (1 mentions)
Ly379268, by Bio-Techne (1 mentions)
Erastin, by Adooq (1 mentions)
Mouse anti α tubulin, by Abcam (1 mentions)
Mouse anti β actin, by Merck Group (1 mentions)
Ecl plus detection system, by GE Healthcare (1 mentions)
Gad67, by Abcam (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
β actin, by Abcam (1 mentions)
6 protocols using horseradish peroxidase conjugated anti mouse and anti rabbit secondary antibodies
1
Glutamate Receptor Signaling Assay
2
Western Blot Analysis of Bacterial Proteins
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SDS-PAGE, transfer of TieA to PVDF membranes and its probing with primary and secondary antibodies were carried out as per standard procedures. For TieA detection, anti-TieA was used at 1:5000 dilutions in 1× PBST (1× PBS + 0.05% Tween 20) for 3 h at room temperature. phospho-ERK, phospho-JNK and phospho-p38 antibodies (eBiosciences, USA) were used at 1:500 dilution in 1× TBST for 3 h. Anti-CagA (Santa Cruz) was used at 1:1000 dilution in 1× TBST. Horseradish peroxidase-conjugated anti-mouse and anti-rabbit secondary antibodies (Sigma-Aldrich) were used at 1:80 000 dilutions. Development of the blots was done using SuperSignal™ West Pico Chemiluminescent Substrate (Thermo Scientific).
3
Radioligand Binding Assay Protocols
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[2,3-3H]D-Asp (specific activity 11.3 Ci/mmol) was from Perkin Elmer (Boston, MA, USA). LY379268 was purchased from Tocris Bioscience (Bristol, UK). (±)-1-(2,5-Dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride ((±)DOI), trazodone, horseradish peroxidase-conjugated anti-mouse, and anti-rabbit secondary antibodies were from Sigma (Milan, Italy). Luminata Forte Western blotting was purchased from Millipore (Temecula, CA, USA). Rabbit anti-GAPDH antibody was from Abcam (Cambridge, UK), rabbit anti-mGlu2/3 antibody was from Novus Biologicals (Littleton CO, USA), rabbit anti–5-HT2A antibody was from Immunostar (Hudson, WI, USA).
4
Transferrin-Mediated Iron Regulation
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Erastin was purchased from AdooQ Bioscience (Cat. No. A13822, Irvine, CA). Ferristatin II (also called chlorazol black) was purchased from Tokyo Chemical Industry Co. (Cat. No. C0533, Tokyo, Japan). Human apo-transferrin (aTf, Cat. No. T2036) and human holo-transferrin (hTf, Cat. No. T0665) were purchased from Sigma-Aldrich (St. Louis, MO). The following antibodies were used for western blotting: rabbit anti-TfR1 (Cat. No. ab84036, Abcam, Cambridge, UK), mouse anti-α-tubulin (Cat. No. T9026, Sigma-Aldrich), mouse anti-β-actin (Cat. No. A3854, Sigma-Aldrich), and horseradish peroxidase-conjugated anti-rabbit and anti-mouse secondary antibodies (Cat. Nos. W401B and W402B, Promega, Madison, WI).
5
Mouse Brain Protein Expression Analysis
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Mice brain were prepared as previously described [63 (link)]. Briefly, mice brain homogenized with RIPA buffer (Thermo Fisher Scientific, Rockford, IL, USA), and incubated on ice for 60 min, and centrifuged at 13,000 rpm for 20 min at 4 °C. An equal amount of total protein (30 μg) was subjected to SDS-PAGE (12%), and the membranes were incubated with the following primary antibodies: CB1 (1:500, Abcam, Cambridge, MA, USA), GAD67 (1:1000, Abcam, Cambridge, MA, USA), β-actin (1:1000, Abcam, Cambridge, MA, USA), and GAPDH (1:500, Cell Signaling Technology, Danvers, MA, USA). The membranes were then incubated with horseradish peroxidase-conjugated anti-rabbit and anti-mouse secondary antibodies (1:5000, Sigma-Aldrich, St. Louis, MO, USA). Immunoreactivity was visualized with an ECL Plus detection system (GE Healthcare, Chicago, IL, USA). The relative density of the protein bands was analyzed with ImageJ.
6
Antibody Characterization for Research
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The following rabbit antibodies were used: Flag (Sigma-Aldrich, St Louis, MO, United States), c-Myc-HRP (Miltenyi Biotec, North Rhine-Westphalia, Germany); RPL11 (D1P5N), Phospho-4EBP1 (Thr70), 4EBP1, S6 ribosomal protein (5G10), Phospho-S6 ribosomal protein (Ser235/236) and TSC2 (Cell Signaling, Danvers, MA, United States) and HA-HRP (Thermo Fisher Scientific, Waltham, Massachusetts, United States). Mouse monoclonal antibodies used were: HA, c-Myc, RPS23 (1E3) and Tubulin (B512) (Sigma-Aldrich, St Louis, MO, United States); eIF4E (BD Biosciences, Franklin Lakes, New Jersey, United States); p72 (1BC11), p72 (18BG3), and p150 (17AH2) (Ingenasa, Madrid, Spain); Cullin4B and RPS23 (Abcam, Cambridge, United Kingdom) and p30 (a gift from J.M. Escribano, Algenex, Madrid, Spain). Horseradish peroxidase-conjugated anti-rabbit and anti-mouse secondary antibodies were from Sigma-Aldrich (St Louis, MO, United States), and Alexa-Fluor−488, −594, and −647 conjugated anti-rabbit and anti-mouse antibodies were from Thermo Fisher Scientific (Waltham, Massachusetts, United States).
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