TXNIP Rabbit Polyclonal Antibody

Flow Cytometry analysis of THP-1 cells using anti-TXNIP antibody. Overlay histogram showing THP-1 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TXNIP Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IHC analysis of TXNIP using anti-TXNIP antibody. TXNIP was detected in paraffin-embedded section of human ovarian cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-TXNIP Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of TXNIP using anti-TXNIP antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: rat brain tissue lysates, Lane 2: rat kidney tissue lysates, Lane 3: rat PC-12 whole cell lysates, Lane 4: mouse brain tissue lysates, Lane 5: mouse kidney tissue lysates, Lane 6: mouse NIH-3T3 whole cell lysates, Lane 7: mouse RAW264.7 whole cell lysates, Lane 8: human K562 whole cell lysates, Lane 9: human Hela whole cell lysates, Lane 10: human HEK293 whole cell lysates, Lane 11: human HL-60 whole cell lysates, Lane 12: human A549 whole cell lysates, Lane 13: human Caco-2 whole cell lysates, Lane 14: human Raji whole cell lysates, After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-TXNIP antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for TXNIP at approximately 44 KD. The expected band size for TXNIP is at 44 KD.