Anti-GPCR RDC1/CXCR-7/ACKR3 Antibody Picoband®

Western blot analysis of GPCR RDC1/CXCR-7/ACKR3 using anti-GPCR RDC1/CXCR-7/ACKR3 antibody (A02656-2).
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 30 ug of sample under reducing conditions.
Lane 1: human RT4 whole cell lysates,
Lane 2: human Hela 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-GPCR RDC1/CXCR-7/ACKR3 antigen affinity purified polyclonal antibody (Catalog # A02656-2) 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 GPCR RDC1/CXCR-7/ACKR3 at approximately 41 kDa. The expected band size for GPCR RDC1/CXCR-7/ACKR3 is at 41 kDa.

LPS induced changes in CXCR7 expression in SGC7901 cells. A: CXCR7 protein expression was assessed by western blotting after SGC7901 cells were stimulated for different periods of time with 500 ng/mL LPS; B: SGC7901 cells were cultured with various concentrations of LPS for 24 h, and CXCR7 protein expression was analyzed via western blotting; C: After exposure of SGC7901 cells to LPS (500 ng/ml), CXCR4 protein expression was assessed by western blotting; D and E: After pretreatment with CCX771, SGC7901 cell proliferation and migration were largely inhibited in response to CXCL12 (100 ng/ml) after 48 h of incubation with LPS. *, P < 0.05, vs. the NC group. The data are presented as the mean ± SD
Index in PubMed under a CC BY license. PMID: 30636642

The effect of TLR4 and MD-2 knockdown on LPS-induced CXCR7 expression. a - c : Gastric cancer cells were transfected with TLR4-specific or MD-2-specific shRNAs, and endogenous TLR4 and MD-2 expression levels were analyzed via qRT-PCR ( a and b ) and western blotting ( c ); d and e : Gastric cancer cells were transfected with TLR4-specific or MD-2-specific shRNAs and treated with 500 ng/mL LPS. A CCK-8 assay was then used to detect cell proliferation ( d ), and a transwell assay was used to assess cell migration ( e ); f and g : As described in D and E, CXCR7 expression was analyzed via RT-PCR (F) and western blotting ( g ) *, P < 0.05, vs. the Lipo2000 group and sh-NC group. The data are presented as the mean ± SD
Index in PubMed under a CC BY license. PMID: 30636642

TLR4, MD-2 and CXCR7 expression in gastric cancer indicates poor prognosis. a : Representative immunohistochemical staining of TLR4, MD-2 and CXCR7 in gastric cancer tissues and paracancerous tissues (original magnification 400×); survival curve for patients with gastric cancer expressing TLR4 ( b ), MD-2 ( c ) and CXCR7 ( d )
Index in PubMed under a CC BY license. PMID: 30636642

LPS enhances the tumorigenicity of SGC7901 cells via the TLR4/MD-2 pathway. a and b : SGC7901 cells (2 × 10 6 cells/mice) treated with different shRNAs (sh-NC, sh-TLR4 and sh-MD-2) were injected subcutaneously into the flanks of nude mice, and the mice were intratumorally injected with LPS (400 μg/kg) every other day. The tumor volume was observed ( a ) and measured ( b ); *, P < 0.05. The data are presented as the mean ± SD; C: CXCR7 expression in tumors was analyzed via immunohistochemistry (original magnification 400×)
Index in PubMed under a CC BY license. PMID: 30636642

IHC analysis of GPCR RDC1/CXCR-7/ACKR3 using anti-GPCR RDC1/CXCR-7/ACKR3 antibody (A02656-2).
GPCR RDC1/CXCR-7/ACKR3 was detected in a paraffin-embedded section of human breast cancer 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 2 μg/ml rabbit anti-GPCR RDC1/CXCR-7/ACKR3 Antibody (A02656-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of GPCR RDC1/CXCR-7/ACKR3 using anti-GPCR RDC1/CXCR-7/ACKR3 antibody (A02656-2).
GPCR RDC1/CXCR-7/ACKR3 was detected in a paraffin-embedded section of human colon adenocarcinoma 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 2 μg/ml rabbit anti-GPCR RDC1/CXCR-7/ACKR3 Antibody (A02656-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of GPCR RDC1/CXCR-7/ACKR3 using anti-GPCR RDC1/CXCR-7/ACKR3 antibody (A02656-2).
GPCR RDC1/CXCR-7/ACKR3 was detected in a paraffin-embedded section of human endometrial cancer 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 2 μg/ml rabbit anti-GPCR RDC1/CXCR-7/ACKR3 Antibody (A02656-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of GPCR RDC1/CXCR-7/ACKR3 using anti-GPCR RDC1/CXCR-7/ACKR3 antibody (A02656-2).
GPCR RDC1/CXCR-7/ACKR3 was detected in a paraffin-embedded section of human urothelial carcinoma 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 2 μg/ml rabbit anti-GPCR RDC1/CXCR-7/ACKR3 Antibody (A02656-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of GPCR RDC1/CXCR-7/ACKR3 using anti-GPCR RDC1/CXCR-7/ACKR3 antibody (A02656-2).
GPCR RDC1/CXCR-7/ACKR3 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 2 μg/ml rabbit anti-GPCR RDC1/CXCR-7/ACKR3 Antibody (A02656-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Flow Cytometry analysis of SiHa cells using anti-GPCR RDC1/CXCR-7/ACKR3 antibody (A02656-2).
Overlay histogram showing SiHa cells stained with A02656-2 (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-GPCR RDC1/CXCR-7/ACKR3 Antibody (A02656-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.