Total cell protein extracts were prepared from PBMCs stimulated with Mtb-Ag for 5 days. In different experiments, PBMCs stimulated with Mtb-Ag for 5 days were exhaustively washed and then cultured in complete RPMI-1640 medium supplemented with 100 U/mL of rhIL-2 for at least 7 days before obtaining total cell protein extracts. Western blotting was performed by standard methods. Each nitrocellulose membrane was blotted with mouse monoclonal antibodies to SH2D1a (SAP, cat. 14-9888-82, eBioscience), stripped, and reblotted with GAPDH (Ambion) or β-actin (BD Biosciences). Bound antibodies were detected with an anti-mouse HRP-conjugated antibody (BioRad, Hercules, CA, USA) using Amersham ECL PLUS reagent (GE Healthcare). Images were obtained with an Intelligent Dark Box (Fujifilm LAS1000) and analyzed with ImageJ Analysis software (NIH, Bethesda, MA, USA). The intensity of each band was expressed as arbitrary units (AU).
Anti mouse hrp conjugated antibody
Manufactured by Bio-Rad
Sourced in United States
Anti-mouse HRP-conjugated antibody is a laboratory reagent used in various immunoassay techniques. It consists of an antibody specific to mouse immunoglobulins that is conjugated to the enzyme Horseradish Peroxidase (HRP). This conjugate allows for the detection and quantification of mouse-derived proteins or antigens in samples.
Automatically generated - may contain errors
Lab products found in correlation
Amersham ecl plus reagent, by GE Healthcare (2 mentions)
Gapdh, by Thermo Fisher Scientific (2 mentions)
β actin, by BD (2 mentions)
Ampicillin sodium salt, by Merck Group (1 mentions)
Polygalacturonic acid sodium salt, by Merck Group (1 mentions)
1 6 diphenyl 1 3 5 hexatriene dph, by Merck Group (1 mentions)
D galacturonic acid monohydrate, by Merck Group (1 mentions)
Iodixanol visipaque, by GE Healthcare (1 mentions)
Hrp conjugated anti flag antibody, by Merck Group (1 mentions)
4 protocols using anti mouse hrp conjugated antibody
1
Western Blot Analysis of SH2D1a Protein
2
Western Blot Analysis of SH2D1a Protein
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Total cell protein extracts were prepared from PBMCs stimulated with Mtb-Ag for 5 days. In different experiments, PBMCs stimulated with Mtb-Ag for 5 days were exhaustively washed and then cultured in complete RPMI-1640 medium supplemented with 100 U/mL of rhIL-2 for at least 7 days before obtaining total cell protein extracts. Western blotting was performed by standard methods. Each nitrocellulose membrane was blotted with mouse monoclonal antibodies to SH2D1a (SAP, cat. 14-9888-82, eBioscience), stripped, and reblotted with GAPDH (Ambion) or β-actin (BD Biosciences). Bound antibodies were detected with an anti-mouse HRP-conjugated antibody (BioRad, Hercules, CA, USA) using Amersham ECL PLUS reagent (GE Healthcare). Images were obtained with an Intelligent Dark Box (Fujifilm LAS1000) and analyzed with ImageJ Analysis software (NIH, Bethesda, MA, USA). The intensity of each band was expressed as arbitrary units (AU).
3
Comprehensive Materials for Biochemical Assays
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All materials were purchased from Pol-Aura (Dywity, Poland) unless otherwise stated, and were of analytical grade. Polygalacturonic acid sodium salt (PGA, from citrus fruit, ≥75%), 1,6-diphenyl-1,3,5-hexatriene (DPH, 98%), D-galacturonic acid (monohydrate), ampicillin sodium salt, and diethyldithiocarbamic acid diethylammonium salt (DIECA, 97%) were purchased from Sigma-Aldrich (Saint Louis, MI, USA). Egg yolk lecithin was obtained from Thermo Fisher Scientific (Waltham, MA, USA). Iodixanol (Visipaque) was purchased from GE Healthcare (Chicago, IL, USA). Lysogeny Agar (LA), Muller-Hinton (MH) broth, and phosphate-buffered saline (PBS) were obtained from Graso Biotech (Owidz, Poland). M63 medium was prepared as described in [44 ]. Crystal Violet Pectate (CVP) medium was prepared according to [32 (link)]. The materials for sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), western blot, and anti-mouse-HRP conjugated antibody were obtained from Bio Rad (Hercules, CA, USA). The anti-gfp antibody (ab 1218) was purchased from Abcam (Cambridge, UK).
4
Flag-SPARTAN Expression Verification
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Flag-SPARTAN expression was verified with Western blot analysis. Cells were collected, lysed, and boiled in 2xSDS-containing buffer for 10 minutes. Electrophoresis was carried out using 10% denaturing polyacrilamide gel. After ON blotting, the expression level was detected using anti-Flag HRPconjugated antibody (Sigma, Cat. No. A8592) diluted 1:5000. As control, we used anti-PCNA HRPconjugated antibody (Santa Cruz, Cat. No. D1811) diluted 1:3000, anti-pol δ antibody (Santa Cruz, Cat. No. J2511) diluted 1:2000, and anti-mouse HRP conjugated antibody diluted 1:5000 (Bio Rad, Cat. No. 170-6516). Detection was carried out using Kodak Imager.
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