Cxcl13
Manufactured by Abcam
Sourced in United States, United Kingdom
CXCL13 is a chemokine that plays a key role in the recruitment and activation of B cells. It is involved in the organization of lymphoid follicles and the migration of B cells to these structures.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710 confocal microscope, by Zeiss (3 mentions)
Ccl21, by Abcam (2 mentions)
Ccl20, by Abcam (1 mentions)
Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)
Cxcr4, by Abcam (1 mentions)
S1pr3, by Abcam (1 mentions)
Ccl25, by Abcam (1 mentions)
Mmp 9, by Abcam (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Huvecs, by Solarbio (1 mentions)
Cxcl13, by Solarbio (1 mentions)
Huvecs, by Selleck Chemicals (1 mentions)
Sb203580, by Selleck Chemicals (1 mentions)
5 protocols using cxcl13
1
Cytokine and Antibody Quantification
2
Kidney Biopsy Immunofluorescence Staining
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Only second kidney biopsy was available for immunofluorescence staining that was adapted from Gu-Trantien et al. [34 (link)]. We used the following primary antibodies: CD20 (ab9475, 1/50), CXCL13 (AF801, 1/200), and CD21 (ab75985, 1/500) and donkey anti-mouse (ab98767, 1/200), anti-goat (ab98514, 1/200), and anti-rabbit (ab98491, 1/200) secondary antibodies. Except for CXCL13 obtained from R&D System (Abingdon, UK), all were provided from Abcam (Cambridge, UK). Briefly, after paraffin removal with xylene, mounted sections were pretreated (citrate 10 mM, pH 6.0, at 95°C to 99°C for 30 minutes), blocked with 1% BSA for 30 min, and incubated with primary antibodies (at 4°C, moist chamber, overnight). After washing, secondary antibodies were applied for 2 h at room temperature. Slides were mounted in the ProLong Gold Antifade Mountant with DAPI (P36941, Life Technologies, California, USA) and images were acquired with a Zeiss LSM 710 confocal microscope.
3
Immune Cell Marker Staining Protocol
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The following monoclonal antibodies were used: CXCR4, CXCL13, CCL21, S1PR3 and αL (Abcam, Cambridge, UK); CXCR5, CXCL12 (R&D Systems, Oxford, UK); CCR7 (Novus Biologicals, Cambridge, UK); S1PR1, α4 (Santa Cruz, Middlesex, UK); S1PR2 (Sigma, Poole, UK). Mouse IgG1 and rabbit Ig were used as negative controls (R&D Systems, Abingdon, UK). All antibodies were titrated on normal tissues and used at saturating concentrations (Additional file 1 : Table S3).
4
Quantifying Immune Chemokines and MMPs in Tumor Microenvironment
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An enzyme-linked immunosorbent assay (ELISA) was used to measure the concentrations of CCL19 (cat. no. ab100601; Abcam, Cambridge, UK), CCL21 (cat. no. ab208985; Abcam), CCL25 (cat. no. ab100645; Abcam), CXCL13 (cat. no. ab179881; Abcam), matrix metalloproteinase (MMP)2 (cat. no. ab100606; Abcam) and MMP9 (cat. no. ab100610; Abcam) in the cell lysate or tumor tissues. The samples from mice were collected after inoculation for 50 days and samples from cells were collected after inoculation for 48 h. All ELISA kits were purchased from Thermo Fisher Scientific, Inc. (eBioscience), and the experiments followed the manufacturer's protocol. The total protein concentration was normalized to the concentration in the samples prior to incubation with vectors or to inoculated animals.
5
HUVEC Inflammation and Permeability Assay
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HUVECs from the American Type Culture Collection (Rockville, MD, USA) were maintained in a humidified incubator at 37 °C, 5% CO2 with Dulbecco’s modified Eagle medium (Hyclone, Logan, UT, USA) containing 10% fetal bovine serum (Gibco, Carlsbad, CA, USA).
HUVECs were exposed to a series of LPS solutions (0, 50, 100, and 200 ng/ml; Solarbio, Beijing, China) for 24 h or treated with 100 ng/ml LPS for 0, 12, 24, and 48 h. The release of CXCL13, as well as the mRNA and protein level of CXCL13/CXCR5, was assessed by enzyme-linked immunosorbent assay (ELISA), real-time PCR, and western blotting, respectively.
HUVECs were transfected with CXCL13 siRNA (siCXCL13#1, 5′-GGUGUUCUGGAGGUCUAUU-3′; siCXCL13#2, 5′-CCAAGAGAGCUCAGUCUUU-3′; and siCXCL13#3, 5′-GGAAGAAGAACAAGUCAAU-3′) or control siRNA (siNC) for 24 h and then treated with 100 ng/ml LPS for 24 h; HUVECs were treated with a p38 inhibitor SB203580 (20 μM; Selleck Chemicals, Houston, TX, USA) or vehicle (DMSO) in the presence of 100 ng/ml CXCL13 (Abcam, Cambridge, MA, USA). Transendothelial electrical resistance (TER) assay and fluorescein isothiocyanate (FITC)-dextran assay were performed to evaluate cell permeability.
HUVECs were exposed to a series of LPS solutions (0, 50, 100, and 200 ng/ml; Solarbio, Beijing, China) for 24 h or treated with 100 ng/ml LPS for 0, 12, 24, and 48 h. The release of CXCL13, as well as the mRNA and protein level of CXCL13/CXCR5, was assessed by enzyme-linked immunosorbent assay (ELISA), real-time PCR, and western blotting, respectively.
HUVECs were transfected with CXCL13 siRNA (siCXCL13#1, 5′-GGUGUUCUGGAGGUCUAUU-3′; siCXCL13#2, 5′-CCAAGAGAGCUCAGUCUUU-3′; and siCXCL13#3, 5′-GGAAGAAGAACAAGUCAAU-3′) or control siRNA (siNC) for 24 h and then treated with 100 ng/ml LPS for 24 h; HUVECs were treated with a p38 inhibitor SB203580 (20 μM; Selleck Chemicals, Houston, TX, USA) or vehicle (DMSO) in the presence of 100 ng/ml CXCL13 (Abcam, Cambridge, MA, USA). Transendothelial electrical resistance (TER) assay and fluorescein isothiocyanate (FITC)-dextran assay were performed to evaluate cell permeability.
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