CD106 (VCAM-1) Monoclonal Antibody (429), eBioscience™
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Figure 3 Loss of endothelial SENP1 inhibits EC activation. ( a ) Grafts from WT or SENP1-ecKO mice were harvested 3 days post-transplantation. The induction of endothelial adhesion molecules was demonstrated by immunofluorescence staining of ICAM-1, VCAM-1, or P-selectin and PECAM-1 with DAPI labelling of the nuclei. Bar represents 50 mum. ( b - e ) Attenuated induction of adhesion molecules in SENP1-ecKO MAECs. Flow cytometry analysis of ICAM-1, VCAM-1 and P-selectin in MAECs isolated from WT or SENP1-ecKO mice after TNF or IL-1beta treatment. Representative histograms are shown in ( b ) with the quantification of mean intensity in ( c - e ). ( f - h ) Overexpression of the catalytically inactive form of SENP1 (SENP1-Mut) inhibits the induction of adhesion molecules in HUVECs. HUVECs were infected by Ad-SENP1-Mut or vector control (Ad-LacZ) for 24 h, treated with pro-inflammatory cytokines and analysed by flow cytometry in the same way as MAECs. Representative histograms of ICAM-1 and VCAM-1 are shown in ( f ) with the quantification of mean intensity in ( g , h ). Data are presented as the mean+-s.e.m. from at least three independent experiments. * P <0.05 and ** P <0.01; two-way ANOVA followed by Bonferroni post-test. MAEC, mouse aortic endothelial cell.
Figure 6 Changes in aortic root and aortic arch mRNA expression for cytokines and adhesion molecules (tumor necrosis factor alpha, chemokine (C-C motif) ligand 2, intercellular adhesion molecule 1 and vascular cell adhesion molecule 1) determined using RT-PCR in wild-type mice. (A) Relative expression at days 1, 3 and 5 days are presented. CCL2, Chemokine (C-C motif) ligand 2; CLP, Cecal ligation and puncture; TNF-alpha, Tumor necrosis factor alpha. Immunofluorescent images of the aortic root obtained to determine the expression of adhesion molecules intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) (left to right: secondary antibody-only control, sham, cecal ligation and puncture (CLP)). ICAM-1 is shown in green, VCAM-1 in red and nuclear material in blue (4',6-diamidino-2-phenylindole dihydrochloride (DAPI)). Inset shows possible colocalization of ICAM-1 with VCAM-1. (B) Graph presents cell adhesion to inflamed aortic endothelium at days 1, 3 and 5 days. Representative images show monocyte adhesion to the endothelium by day 5 (* P < 0.05 compared to the time-matched sham-treated groups).
Figure 1 Low doses of ionizing radiation have no significant impact on cell death and migration properties of FLS cultures of healthy mice. Healthy FLS cultures obtained from C57Bl/6 mice were phenotyped at passage 5, seeded and analyzed 96 h after the treatment with low, intermediate, and high doses of ionizing radiation. Cell pools that were tested via flow cytometry were considered to be FLS when being CD11b - ( A.1 ), CD54 + ( A.2 ), and CD106 + ( A.3 ). B.1 to B.3 shows gating strategy for cell death analysis of FLS via flow cytometry. Cell death was determined after staining of the cells ( B.1 ., total scatter) with AxV-FITC/PI ( B.2 , B.3 ). Vital cells were defined as AxV - /PI - , necrotic ones as PI + ( B.2 , E ), and apoptotic cells as AxV + , but PI - ( B.3 , D ). Cell numbers ( C ) of living FLS were counted using a Neubauer chamber. Cell migration and invasiveness was tested via scratch assay ( F ) and matrigel invasion assay ( G ). For the scratch assay, cells were irradiated after a scratch was applied; pictures were taken every 24 h and scratch area was quantified using Tscratch (V1.0, Copyright T.Geback, M.Schulz, Zurich, Switzerland). The initial scratch area was set to 100% and scratch area of the consecutive days was calculated accordingly. For assessment of invasiveness cells were seeded in a matrigel coated transwell, irradiated and incubated for 3 days. Cells were then stained with crystal violet and invaded area was assessed using Image
Fig. 3 The phosphorylation level of JNK/MAPK signaling pathway are downregulated during developing mouse embryos muscle and primary skeletal muscle cell differentiation. a The protein expression levels of myogenic markers (MyoD and MyoG) in developing mouse embryos muscle. Mouse hind limb muscles were isolated from seven time points: E12.5, E15.5, E18.5, postnatal 1 week, postnatal 2 weeks, postnatal 4 weeks, and postnatal 8 weeks (adult). b Western blotting for phosphorylation level of JNK/MAPK signaling: JNK, c-JUN, and their phosphorylated forms p-JNK and p-c-JUN in developing mouse embryos muscle. c Relative expression in a and b were calculated. Data in the same group were compared to E12.5. GAPDH was the internal control. d The protein expression levels of myogenic markers (MyoD and MyoG) during primary skeletal muscle cell differentiation. MuSCs were isolated from hind limb muscles in postnatal 1-week mice, and then the cells were activated in vitro for expansion of myogenic progenitors (D0) and subsequently to differentiation (D1 to D8). e Western blotting for phosphorylation level of JNK/MAPK signaling: JNK, c-JUN, and their phosphorylated forms p-JNK and p-c-JUN during primary skeletal muscle cell differentiation. f Relative expression in d and e were calculated. Data in the same group were compared to D0 (except for MyoD was compared to D2). GAPDH was the internal control. Values are means +- SEM. The statistical significance of difference betwe
Figure 8 Treg transmigration through lymphatic endothelium depends on VCAM-1 in vitro and in vivo . ( a ) SVEC4-10 or mouse skin LEC: grey histograms, isotype control; blue: LTbetaR. Representative of 3 experiments. ( b ) SVEC4-10 cultured 48 h, with or without agonistic 1 mug ml -1 anti-LTbetaR last 24 h, stained for indicated molecules. Isotype control, black histograms; indicated antibody, blue (control-treated) and red (anti-LTbetaR-treated). Representative of two experiments. ( c , d ) Aza Treg migrated across iSVEC4-10 to CCL19. SVEC4-10 (anti-ICAM-1 and anti-VCAM-1) or Treg (anti-LFA-1, anti-VLA-4, MOPC21 and LTbetaRIg) pretreated as indicated. Migration normalized to control. Six transwells from two experiments in c and nine transwells from three experiments in d . ( e ) Footpad migration. CFSE-labelled WT nTreg or naive CD4+ non-Treg co-injected with Rat IgG2a or anti-VCAM-1 or pretreated with Rat IgG2b or anti-VLA-4. Per cent CFSE+ cells of popliteal LN CD4 T cells shown. Treg: 5-8 mice from 3 experiments; non-Treg: 7-10 mice from 3 experiments. ( f - i ) Aza Treg transwell migration across iSVEC4-10 ( f , g ) and primary mouse iLEC ( h , i ). Under static conditions ( f , h ) and with fluid flow ( g , i ). T cells (MOPC21 and LTbetaRIg) or endothelial cells (NIKi and anti-VCAM-1) treated as indicated. White numerals indicate % decline compared with MOPC21. Migration normalized to MOPC21. Results from nine transwells per condition from three experiments in f and g ,
Published figure using CD106 (VCAM-1) monoclonal antibody (Product # 14-1061-82) in Flow Cytometry
Description: The 429 monoclonal antibody reacts with mouse CD106 (Vascular Cell Adhesion Molecule-1, VCAM-1), a 110 kDa transmembrane glycoprotein expressed by myeloid lineage and bone marrow stromal cells. Endothelial cells constitutively express low levels of CD106 and upregulate it upon cytokine stimulation. CD106 binds to integrin alpha4beta1 (VLA-4, CD49d/CD29) and Integrin a4b7 (LPAM-1) and these interactions in the bone marrow and thymus are important for early lymphocyte and myeloid development. Cytokine-mediated upregulation of CD106 on endothelial cells suggests a role for this antigen in the inflammatory response.
Applications Reported: This 429 antibody has been reported for use in flow cytometric analysis, immunoprecipitation, and immunohistochemical staining of frozen tissue sections (IHC-F). It has also been reported in blocking of CD106 in functional studies. (Please use Functional Grade purified 429, cat. 16-1061, in functional assays.). Applications Tested: The 429 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This can be used at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
VCAM-1 is a cell surface sialoglycoprotein expressed by cytokine activated endothelium. The protein has a number of functions including the regulation of leukocyte migration, leukocyte-endothelial cell adhesion and signal transduction and may play a role in a number of inflammatory diseases.
DKFZp779G2333; INCAM100; MGC99561; sCD106; soluble CD106; soluble VCAM 1; sVCAM 1; V-CAM 1; vascular cell adhesion protein 1
Entrez Gene ID: