Anti-CXCR2 Antibody Picoband®

Western blot analysis of CXCR2 using anti-CXCR2 antibody (A00455-2).
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 30ug of sample under reducing conditions.
Lane 1: human U20S whole cell lysates,
Lane 2: human A549 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-CXCR2 antigen affinity purified polyclonal antibody (Catalog # A00455-2) 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 CXCR2 at approximately 41KD. The expected band size for CXCR2 is at 41KD.

RM-1 cell inoculation increases CXCR2 mRNA and protein expression in the spinal cord. (A) Real-time PCR results show the increase of CXCR2 mRNA expression in the spinal cord. CXCR2 mRNA was increased from 7 days to 21 days after inoculation. *P <0.05 vs . sham. n = 4 mice per group. (B) Western blot shows time course of CXCR2 protein expression in the spinal cord after inoculation (B) . n = 3 mice per group. (C, D) Immunostaining shows the CXCR2 expression in the spinal cord in sham (C) and inoculated (D) animals. CXCR2-IR was increased at 7 days after inoculation. (E-G) Double staining shows CXCR2 was colocalized with neuronal marker NeuN. H , I . SB2205002 attenuated RM-1 cell inoculation-induced mechanical allodynia (H) and heat hyperalgesia (I) . * P <0.05; ** P <0.01; *** P <0.001 vs . vehicle.
Index in PubMed under a CC BY license. PMID: 24580964

IHC analysis of CXCR2 using anti-CXCR2 antibody (A00455-2).
CXCR2 was detected in paraffin-embedded section of human appendicitis 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 2μg/ml rabbit anti-CXCR2 Antibody (A00455-2) 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 CXCR2 using anti-CXCR2 antibody (A00455-2).
CXCR2 was detected in paraffin-embedded section of human tonsil 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 2μg/ml rabbit anti-CXCR2 Antibody (A00455-2) 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 THP-1 cells using anti-CXCR2 antibody (A00455-2).
Overlay histogram showing THP-1 cells stained with A00455-2 (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-CXCR2 Antibody (A00455-2, 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.