Proteins extracted from L6-S1 spinal cord PAR4- or scramble-treated mice were separated using a 4 – 15% Mini-PROTEAN TGX precast polyacrylamide gel (Bio-Rad, Hercules, CA). After electrophoresis, separated proteins were transferred to a polyvinylidene difluoride membrane. MIF and HMGB1 protein bands were visualized using a biotinylated goat polyclonal primary antibody (BAF289; R&D, Minneapolis, MN; 1:1000) and a rabbit polyclonal primary antibody (ab18256; Abcam, Cambridge, MA; 1:4000), along with GAPDH as an internal control (A00915, GenScript, Piscataway, NJ; 1:2500), a biotinylated anti-rabbit secondary antibody (Vector Labs, Burlingame, CA; 1:400), streptavidin-HRP conjugates and chemiluminescent substrate (Pierce, Rockford, IL). Band densitometry was performed using ImageJ (NIH, Bethesda, MD).
A00915
Manufactured by GenScript
A00915 is a precision electronic balance that can measure weights up to 220 grams with an accuracy of 0.0001 grams. It features a backlit LCD display and is designed for laboratory and research applications.
Automatically generated - may contain errors
2 protocols using a00915
1
Quantifying MIF and HMGB1 in Spinal Cord
2
HMGB1 Quantification in Mouse Bladder
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Proteins extracted from bladder tissue of naïve wild type and MIF knockout mice were separated using a 4–15% Mini‐PROTEAN TGX precast polyacrylamide gel (Bio‐Rad, Hercules, CA). After electrophoresis, separated proteins were transferred to a polyvinylidene difluoride membrane. HMGB1 protein bands were visualized using a rabbit polyclonal primary antibody (ab18256; Abcam, Cambridge, MA; 1:4000), along with GAPDH as an internal control (A00915, GenScript, Piscataway, NJ; 1:2500), a biotinylated anti‐rabbit secondary antibody (Vector Labs, Burlingame, CA; 1:400), streptavidin‐HRP conjugates and chemiluminescent substrate (Pierce, Rockford, IL). Band densitometry was performed using ImageJ (NIH, Bethesda, MD).
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