14-0549-82

antibody from Invitrogen Antibodies

Targeting: ICAM1 BB2, CD54

Provider product page for 14-0549-82

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Antibody data

Antibody Data
Antigen structure
References [16]
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Validations
Other assay [8]

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Product number: 14-0549-82 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD54 (ICAM-1) Monoclonal Antibody (HA58), eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The HA58 monoclonal antibody reacts with human CD54 (InterCellular Adhesion Molecule-1, ICAM-1), a 90-110 kDa transmembrane glycoprotein expressed by monocytes, lymphocytes and endothelial cells. Expression of CD54 is upregulated on activated lymphocytes. Interaction of CD54 with its ligand CD11a is important in the inflammatory response.
Antibody clone number: HA58
Concentration: 0.5 mg/mL

ICAM1 protein structure - 14-0549-82 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 Clinical Relevance of CD54 Expression. ( a ) Relative CD54 mRNA expression in primary (n=24) and recurrent tumors (n=12) from prostate cancer patients. ( b ) Relative CD54 mRNA expression correlated with mortality of patients with prostate cancer by Kaplan-Meier survival analysis. ( c ) CD54 expression in primary and recurrent tumors from two prostate cancer patients. ( d ) Immunohistochemical analysis of in situ CD54 expression in primary and recurrent tumors from two prostate cancer patients. ( e ) Representative light micrograph of the sphere-forming capacity of CD54 + and CD54 - sorted cells isolated from primary and secondary xenograft tumors of prostate cancer patients and quantification of the results. ( f ) Tumor-initiating capacity of differing numbers of different quadrants of CD54 + and CD54 - tumor cells isolated from primary and secondary prostate cancer patient xenograft tumors of NOD/SCID mice 6 weeks after injection. Red numbers indicate number of cells injected, and red arrows indicate tumor formation. ( g ) Immunohistochemical analysis of in situ CD54 expression in both primary and secondary tumor samples collected from xenografted CD54 + tumors. H&E stained tissues are shown as a reference (scale bar=50 mum). ** P < 0.01.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8 GATA2 SUMOylation inhibits its DNA binding activity. ( a ) SUMO conjugation reduces GATA2 DNA binding activity. GATA2-WT, GATA2-2KR, SUMO-GATA2 or control constructs were transfected into 293 T cells, and nuclear extracts were processed with EMSA using a GATA2-specific oligonucleotide probe (top). Input of GATA2 was detected by western blotting with anti-Flag antibody (bottom). ( b ) Overexpression of catalytic inactive form of SENP1 (SENP1-Mut) inhibits recruitment of GATA2 to the promoter of ICAM-1, VCAM-1 and E-selectin. HUVECs were infected by Ad-SENP1-Mut or Ad-LacZ and then treated with TNF. Nuclear extracts were then subjected to ChIP assay with the anti-GATA2 antibody followed by quantitative real-time PCR for the promoter sequences of ICAM-1, VCAM-1 and E-selectin containing a GATA2 binding site. Quantitative results are shown as the ratio of ChIP to input values. Data are presented as the mean+-s.e.m. from three independent experiments. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 9 The critical role of GATA2 SUMOylation in EC activation. ( a ) GATA2 SUMOylation regulates the expression of endothelial adhesion molecules. HUVECs were transfected by GATA2 siRNA or control siRNA for 48 h followed by infection with Ad-GATA2-WT, Ad-GATA2-KR, Ad-SUMO-GATA2 or their vector control (GFP) as indicated. HUVECs were treated with TNF or vehicle control for 24 h, and the cell lysates were then subjected to western blotting with anti-ICAM-1, anti-VCAM-1, anti-E-selectin or anti-GATA2 antibodies. Actin was used as a loading control. ( b ) GATA2 SUMOylation site mutation increases endothelial adhesion molecule induction by TNF at the early phase. The expression of endothelial adhesion molecules was determined by quantitative real-time PCR in HUVECs with GATA2-WT or GATA2-2KR reconstitution as described in ( a ). Data are presented as the mean+-s.e.m. from at least three independent experiments. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 1 CD31 interactions promote the recovery of endothelial integrity following endothelial contraction induced by MHC molecule triggering. a - d Following MHC or ICAM-1 and/or CD31 antibody-mediated co-ligation for 30 min, EC were fixed and stained with rhodamine-phalloidin. Images taken on EC monolayers seeded at identical density are shown in ( a , b ). The average F-actin intensity per cell of three independent experiments is shown in ( c , d ). Scale bar, 20 mum. ( n = 3 biologically independent samples, N = 3 independent experiments, data are mean +- SD). One-way Anova with Tuckey post-hoc test. MHC vs all **** p < 0.0001, MHC + CD31 vs Isc **** p < 0.0001, MHC + CD31 vs all ****p < 0.0001. e Western blot (WB) analysis of Erk activation by WT and cd31 -/- EC 30 min after MHC stimulation. The bar graph shows relative protein expression +- SEM. N = 3 independent experiments (data are mean +- SD). One-way Anova with Tuckey post-hoc test. cd31 -/- MHC vs cd31 -/- IsC *** p = 0.0002, cd31 -/- MHC vs all **** p < 0.0001. f Western blot (WB) analysis of RhoA activation by WT and cd31 -/- EC 30 min after MHC stimulation. The bar graph shows relative protein expression +- SEM. N = 3 independent experiments (data are mean +- SD). One-way Anova with Tuckey post-hoc test. cd31 -/- 15' vs all *** p = 0.0003, cd31 -/- 30' vs all **** p < 0.0001. g Immunoprecipitation of CD31 molecules from WT EC exposed to MHC/ICAM-1 stimulation for 30 min followed by immunoblotting with an anti-pho

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Effect of nicotine on the P. gingivalis LPS-induced protein expression of pro-inflammatory mediators in HUVECs. HUVECs were stimulated by P. gingivalis LPS in the presence or absence of nicotine (10 uM-10 mM) for 4, 24, and 72 h. After stimulation, the surface expression levels of ICAM-1 (A), VCAM-1 (B), and E-selectin (C) were measured by flow cytometry, and the quantity of MCP-1 (D) and IL-8 (E) in conditioned media was measured by ELISA. Each value represents mean +-SD of three independent assays. Non-stimulated HUVECs were used as a control. The protein expression levels of pro-inflammatory mediators were not analyzed after stimulation with 10-mM nicotine for 24 and 72 h because the cells were not viable. * - significantly different between groups, p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Cancer Stem Cell Traits of Differential Populations of CD54 + and CD54 - Cells. ( a ) FACS histograms show separate gating for the isolation of CD54 + and CD54 - PC3 and LNCaP cells. ( b ) Representative light micrograph fields and comparative quantification of the sphere-forming capacity of CD54 + and CD54 - LNCaP and PC3 cells. ( c ) Representative microscope fields from colony formation assays for CD54 + and CD54 - PC3 cells. ( d ) The cisplatin (DDP) IC 50 of CD54 + and CD54 - cells from the PC3, LNCaP, DU145, and 22RV1 prostate cancer cell lines. ( e ) q-PCR analysis of the expression of Hedgehog, Notch, and Wnt signaling pathway genes as well as genes involved in stem cell self-renewal in CD54 + and CD54 - PC3 cells. ( f ) Representative micrographs of in situ protein expression of the key signaling pathway components HES1 and CD54 in DAPI-stained LNCaP and PC3 cells. ** P < 0.01.