Anti-CD45/PTPRC Antibody Picoband®

Effects of long-term L -serine administration on orexigenic neuropeptide expression in aging mice. NPY (A–E) and AGRP (F–J) expression as determined by immunohistochemistry; relative NPY (K) and AGRP (L) mRNA expression. Aged, untreated control mice; Low-ser, mice supplemented with 0.1% (wt/vol) L -serine dissolved in the drinking water; Middle-ser, mice supplemented with 0.2% (wt/vol) L -serine dissolved in the drinking water; High-ser, mice supplemented with 0.5% (wt/vol) L -serine dissolved in the drinking water. Young, adult male mice at the age of 18 months. Arrow (yellow), expression of target proteins; NPY, neuropeptide Y; AGRP, agouti-related protein. Values are expressed as mean ± SEM, n = 8; a, b Means of the bars with different letters were significantly different among groups ( P < 0.05).
Index in PubMed under a CC BY license. PMID: 30190704

Western blot analysis of CD45 using anti-CD45 antibody (PB9096).
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 30 ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: 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-CD45 antigen affinity purified polyclonal antibody (Catalog # PB9096) 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 CD45 at approximately 180-250 kDa. The expected band size for AFP is at 147 kDa.

Flow Cytometry analysis of THP-1 cells using anti-CD45 antibody (PB9096).
Overlay histogram showing THP-1 cells stained with PB9096 (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-CD45 Antibody (PB9096,1μg/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.

IHC analysis of CD45 using anti-CD45 antibody (PB9096).
CD45 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-CD45 Antibody (PB9096) 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.

IF analysis of CD45 using anti-CD45 antibody (PB9096)
CD45 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-CD45 Antibody (PB9096) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of CD45 using anti-CD45 antibody (PB9096)
CD45 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-CD45 Antibody (PB9096) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.