In brief, cultured cells and brain tissues were homogenized in RIRA buffer supplemented with protease and phosphatase inhibitors on ice. The homogenates were centrifuged at 12,000 rpm for 20 min at 4°C. The supernatants were collected and quantified using the bicinchoninic acid (BCA) assay (Pierce Biotechnology). The samples were mixed with loading buffer and boiled for 5 min. Equal amounts of proteins were loaded to each well and separated by SDS-PAGE. Primary antibodies were used at the following dilutions: rabbit anti-TIGAR (1:500, Abcam), mouse anti-Bcl-2 (1:500, Santa), rabbit anti-Bax (1:1,000, CST), rabbit anti-LC3B (1:1,000, CST), rabbit anti-Beclin-1 (1:500, CST), rabbit anti-Caspase-3 (1:1,000, CST), rabbit anti-cleaved-Caspase-3 (1:500, CST), rabbit anti-β-actin (1:2,000, CST), rabbit anti-P62 (1:1,000, CST), rabbit anti-nitric oxide synthase 1 (NOS1; 1:100, CST) and rabbit anti-G6PD (1:1,000, CST).
Rabbit anti tigar
Manufactured by Abcam
Sourced in United Kingdom
Rabbit anti-TIGAR is a primary antibody that specifically binds to the TIGAR (TP53-Induced Glycolysis and Apoptosis Regulator) protein. TIGAR is a regulator of glycolysis and apoptosis. This antibody can be used for the detection and analysis of TIGAR in various research applications.
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Lab products found in correlation
2 protocols using rabbit anti tigar
1
Analysis of Cell Signaling Pathways
2
Quantifying TIGAR and LC3B in tumors
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Immunohistochemical staining was performed with rabbit anti-TIGAR (dilution, 1:400; Abcam, Cambridge, UK, ab37910) and mouse anti-LC3B (dilution 1:200; Cell Signaling Technology, Inc., Danvers, MA, USA, #3868) antibodies. In the negative control group, the primary antibody was replaced by PBS. Each sample was graded according to the intensity and extent of staining. The intensity of staining was scored as 0 (no staining), 1 (weak staining), 2 (warm staining), or 3 (strong staining). The percentage of positively stained tumor cells was scored as 0 (0%), 1 (1%–25%), 2 (26%–50%), 3 (51%–75%), or 4 (76%–100%). The intensity score × the percentage score was used to obtain the final overall score, and the specimens were classified into two groups according to overall score. Samples were considered LC3B− with a score of 0–5 or LC3B+ with a score of 6–12; samples were considered TIGAR− with a score of 0–5, or TIGAR+ with a score of 6–12. All slides were assessed independently by two pathologists blinded to patient identity and clinical outcome.
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