Liver tissue samples taken from the body whose postmortem intervals were within 24 h for western blot analysis. Liver tissues (30 mg) were washed in PBS and homogenized in RIPA buffer (Nacalai Tesque) containing proteinase inhibitor cocktail (Nacalai tesque). Protein was separated on a 10% polyacrylamide gel and transferred electrophoretically to an Immobilon-E transfer membrane (Merck Millipore). Blots were blocked in 1% bovine serum albumin in tris-buffered saline containing 0.1% Tween 20 (TBS-T) for 30 min at room temperature. SMOX, SAT, AcPAO, ODC, SPDS, SPMS, AMD1 and GAPDH were detected by an ECL Western Blotting Detection System (GE Healthcare) using anti-SMOX (Proteintech), anti-SAT (abcam), anti-AcPAO (abcam), anti-ODC (Proteintech), anti-SPDS (Proteintech), SPMS (Proteintech), AMD1 (Proteintech) and anti-GAPDH (Sigma) antibodies. The images were captured using Amersham Imager 600 (GE Healthcare) and the bands were quantified using the ImageJ program (Schneider, C. A.; Rasband, W. S. & Eliceiri, K. W. (2012), "NIH Image to ImageJ: 25 years of image analysis," Nature methods 9 (7): 671–675), normalized to GAPDH and expressed as relative amount.
Anti spds
Manufactured by Proteintech
Anti-SPDS is a laboratory equipment product that is used to detect and analyze the spermidine synthase (SPDS) protein. It functions as a tool for researchers to study the expression and activity of SPDS, which is an enzyme involved in the biosynthesis of polyamines. The product provides a reliable and specific means to measure SPDS levels in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti smox, by Proteintech (2 mentions)
Anti acpao, by Abcam (2 mentions)
Anti odc, by Proteintech (2 mentions)
Proteinase inhibitor cocktail, by Nacalai Tesque (1 mentions)
Ripa buffer, by Nacalai Tesque (1 mentions)
Ecl western blotting detection system, by GE Healthcare (1 mentions)
Amersham imager 600, by GE Healthcare (1 mentions)
Anti gapdh, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Nacalai Tesque (1 mentions)
Anti p21, by Proteintech (1 mentions)
Anti p16, by Proteintech (1 mentions)
Imagequant las 500, by GE Healthcare (1 mentions)
Blocking one reagent, by Nacalai Tesque (1 mentions)
2 protocols using anti spds
1
Western Blot Analysis of Liver Proteins
2
Western Blot Analysis of Polyamine Metabolic Enzymes
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The cells were washed with PBS and homogenized in RIPA buffer containing a protease inhibitor cocktail (Nacalai Tesque, Japan). Proteins were separated on a 10% polyacrylamide gel and electrophoretically transferred to an immunoblotting polyvinylidene difluoride (PVDF) membrane (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The blots were blocked with the Blocking One reagent (Nacalai Tesque) for 30 min at room temperature. SMOX, ODC, AMD1, SPDS, SPMS, SAT1, AcPAO, protein-conjugated acrolein, p21, p16, GAPDH and actin were detected with a Chemi-Lumi One reagent (Nacalai tesque) using anti-SMOX (Proteintech), anti-ODC (Proteintech), anti-AMD1 (Proteintech), anti-SPDS (Proteintech), anti-SPMS (Proteintech), anti-SAT1 (abcam), anti-AcPAO (abcam), anti-protein-conjugated acrolein (abcam), anti-p21 (Proteintech), anti-p16 (Proteintech), anti-GAPDH (invitorgen) and anti-β-actin (MBL) antibodies. Images were acquired using an ImageQuant LAS 500 (GE Healthcare). The bands were quantified using the ImageJ program [54 (link)], normalized to Actin and expressed as relative amount of young, non-treated cells.
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