Anti-CCXCR1/XCR1 Antibody Picoband®

Western blot analysis of CCXCR1/XCR1 using anti-CCXCR1/XCR1 antibody (A04185-1).
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 U-87MG whole cell lysates,
Lane 3: human HEL whole cell lysates,
Lane 4: human U20S whole cell lysates,
Lane 5: rat spleen tissue lysates,
Lane 6: mouse spleen tissue lysates,
Lane 7: mouse thymus tissue 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-CCXCR1/XCR1 antigen affinity purified polyclonal antibody (Catalog # A04185-1) 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 CCXCR1/XCR1 at approximately 42 kDa. The expected band size for CCXCR1/XCR1 is at 39 kDa.

cDC1s in SCI patients and mouse model of SCI. A PBMCs were collected from 10 healthy volunteers and 10 SCI patients. Identification of cDC1s and cDC2s subsets was performed by flow cytometry. CD11C + HLA-DR + represents cDCs, while CD141 + represents sDC1s and CD1c + represents cDC2 cells. Difference was calculated by Mann–Whitney U test. ** P < 0.01 vs. healthy group. B Mice were divided into sham ( n = 6) and SCI ( n = 6) groups. BMS score detection was performed. Difference was calculated by two-way ANOVA test. ** P < 0.01 vs. sham group. C–D The positive expressions of MHCII and XCR1 in spinal cord tissues were detected by IF assay. The difference was calculated by unpaired two-tailed T-test. ** P < 0.01 vs. sham group. SCI : spinal cord injury; dpi : days post-injury; cDC : conventional dendritic cell
Index in PubMed under a CC BY license. PMID: 39901071

cDC1 depletion reduces CD8 + T cell infiltration after SCI injury. A Mice were divided into SCI ( n = 6) and SCI + Qu ( n = 6) groups. Qu administration was started on day 14 after surgery. B BMS score was determined. The difference was calculated by unpaired two-tailed T-test. ** P < 0.01 vs. SCI group. C Positive expression of XCR1 in spinal cord tissues was detected by IF assay. The difference was calculated by unpaired two-tailed T-test. ** P < 0.01. D The results of flow cytometry of XCR1 + cDCs in lymph nodes and blood. The cDC population was expressed as CD11C and I-A/I-E (MHCII) double positive, and XCR1 + represents XCR1 + cDCs. E The proportion of CD8 + T cells in spinal cord tissues was determined by flow cytometry. CD3 + T cells were circled with CD3 gate, and further CD3 + CD8 + T cells were analyzed. The difference was calculated by unpaired two-tailed T-test. ** P < 0.01 vs. SCI group. N = 6. F ELISA was used to detect IFN-γ levels in spinal cord tissues. The difference was calculated by unpaired two-tailed T-test. ** P < 0.01 vs. SCI group. N = 6. dpi : days post-injury; Qu : Quizartinib
Index in PubMed under a CC BY license. PMID: 39901071

Excess cDC1s in non-lymphoid tissues in LNs of SCI mice is due to migration of cDC1s from the spinal cord. A Leukocyte (CD45 + ) and pre-DC subsets in bone marrow of sham and SCI groups were detected by flow cytometry. Immune cells were circled by CD45 + and murine pre-DCs were further detected by CD135 + CD11c + . The difference was calculated by unpaired two-tailed T-test. N = 6. B–C Non-lymphoid tissue cDC1 (CD11C + I-A/I-E + CD103 + ) proportion and lymphoid-resident cDC1 (CD11C + I-A/I-E + CD8α + ) subsets in dcLN and LLN of sham and SCI groups were detected by flow cytometry. The difference was calculated by unpaired two-tailed T-test. ** P < 0.01 vs. SCI group. N = 3. D cDC1s (CD11C + I-A/I-E + CD103 + ) and cDC2s (CD11C + I-A/I-E + CD11b + ) subsets in the blood of SCI-3 dpi and SCI-42 dpi groups were detected by flow cytometry. The difference was calculated by unpaired two-tailed T-test. ** P < 0.01 vs. SCI group. N = 6. E cDC1 subsets (CD45 + XCR1 + CD103 + ) in spinal cord tissues of SCI-3 dpi and SCI-42 dpi groups were detected by flow cytometry. The difference was calculated by unpaired two-tailed T-test. ** P < 0.01 vs. SCI group. N = 6. F The positive expressions of CD11C and LYVE-1 in the spinal cord tissues of SCI group mice were detected by IF assay. BM : bone marrow; dcLN : deep cervical lymph nodes; LLN : lumbar lymph nodes
Index in PubMed under a CC BY license. PMID: 39901071

IHC analysis of CCXCR1/XCR1 using anti-CCXCR1/XCR1 antibody (A04185-1).
CCXCR1/XCR1 was detected in a paraffin-embedded section of human spleen 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-CCXCR1/XCR1 Antibody (A04185-1) 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 CCXCR1/XCR1 using anti-CCXCR1/XCR1 antibody (A04185-1).
CCXCR1/XCR1 was detected in a paraffin-embedded section of mouse thymus 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-CCXCR1/XCR1 Antibody (A04185-1) 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 CCXCR1/XCR1 using anti-CCXCR1/XCR1 antibody (A04185-1).
CCXCR1/XCR1 was detected in a paraffin-embedded section of rat thymus 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-CCXCR1/XCR1 Antibody (A04185-1) 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.