Anti-XBP1 Antibody Picoband®

Western blot analysis of XBP using anti-XBP antibody (PB9463).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: MCF-7 Whole Cell Lysate at 40ug,
Lane 2: MM231 Whole Cell Lysate at 40ug,
Lane 3: MM453 Whole Cell Lysate at 40ug,
Lane 4: SKOV Whole Cell Lysate at 40ug,
Lane 5: HELA Whole Cell Lysate at 40ug.
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-XBP antigen affinity purified polyclonal antibody (Catalog # PB9463) 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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for XBP at approximately 29 kDa. The expected band size for XBP is at 29 kDa.

IHC analysis of XBP using anti-XBP antibody (PB9463).
XBP was detected in a paraffin-embedded section of mouse lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-XBP Antibody (PB9463) 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) (Catalog # SA1022) with DAB as the chromogen.

Inhibition of autophagic flux alleviated hepatocyte ER stress and LD accumulation. (A) Changes in ER morphology and LD accumulation in different groups were assessed. (B) Hepatic XBP1, IRE1α, and EIF2α protein levels were compared among groups. (C) XBP1 and IRE1α protein levels were normalized based on β-Actin levels, and the p-EIF2α/EIF2α ratio was compared among groups. (Yellow arrow: LDs; black arrow: ER; * p < 0.05, ** p < 0.01, ns: no statistical difference).
Index in PubMed under a CC BY license. PMID: 35982710

IHC analysis of XBP using anti-XBP antibody (PB9463).
XBP was detected in a paraffin-embedded section of rat lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-XBP Antibody (PB9463) 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) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of XBP using anti-XBP antibody (PB9463).
XBP was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-XBP Antibody (PB9463) 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) (Catalog # SA1022) with DAB as the chromogen.

Flow Cytometry analysis of HepG2 cells using anti-XBP1 antibody (PB9463).
Overlay histogram showing HepG2 cells stained with PB9463 (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-XBP1 Antibody (PB9463, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.