monoclonal antihuman resistin  (R&D Systems)

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 1. Immunohistochemical staining of aneurysms with antibodies to resistin and CD68. The various types of human vessels were stained with antibodies against resistin or CD68, the latter of which is a marker of monocytes/macrophages. Mayer’s hematoxylin was used for counterstaining. The left upper region of each picture indicates the endothelial lining. Resistin-positive areas are seen in atherosclerotic lesions as dark staining beneath the endothelial lining (C and D). Resistin-positive areas are distributed along inflammatory cells and stained positively for CD68 (G, H). No immunoreactive area was observed for either of the two antibodies in normal vessels (A, B, E, F). Staining without primary Abs was performed as a negative control (I, J, K, L).

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Immunohistochemical staining, Staining, Marker, Negative Control

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 3. Resistin mRNA expression measured by real-time PCR. (A) Resistin mRNA expression in aortic aneurysms. Total RNA was purified from aneurysm centers (n =4) and veins (n =3). Real-time quantitative PCR revealed the overexpression of resistin in aneurysm centers. The y axis represents resistin expression normalized against GAPDH expression. The data illustrated on the graph represent meansTSEM of 2deltadeltaCt of the samples. (B) Resistin mRNA expression in vascular cells. Isolated human VSMCs and commercial HUVECs were incubated with 10 ng/mL of TNF-a for 12 h. Human monocytes were isolated and incubated for 36 h, and total RNA was purified and resistin mRNA levels were measured by real-time RT-PCR, normalized with respect to the GAPDH mRNA levels. The data illustrated on the graph represent meansTSEM of 2deltadeltaCt of three different experiments.

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Expressing, Real-time Polymerase Chain Reaction, Purification, Over Expression, Isolation, Incubation, Quantitative RT-PCR

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 2. Immunofluorescent double staining of aneurysms with antibodies to resistin and vascular cell markers. Green indicates vascular cell proteins, red resistin, and blue DAPI-stained cellular nuclei. The upper region in each picture indicates endothelial lining. Resistin protein was found to co-localize with CD68 (A, B). Smooth muscle (SM) actin was positive throughout the vessel thickness (D), whereas CD31 was not detected (G). The distribution of resistin (E, H) was not concordant with either D or G. A, B and C were magnified (J¨L) and this showed that resistin was present both intracellularly and extracellularly. The merged picture (M) shows yellow rings composed of red resistin-positive areas and green transmembrane CD68-positive areas (arrows).

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Double Staining, Staining

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 4. Induction of PAI-1 in HUVECs after resistin treatment. HUVECs were incubated with 10¨100 ng/mL of resistin for 4¨24 h or with 10 ng/mL of TNF- a for 12 h as a positive control. (A) PAI-1 protein levels in culture media were measured by ELISA and are presented as fold changes versus the negative control. (B) PAI-1 mRNA expression was analyzed by Northern blotting, and normalized against GAPDH mRNA. Results are presented as fold changes versus the negative control. The data illustrated on the graph represent the meansTSEM of three different experiments. *p <0.05 vs. without resistin treatment.

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Incubation, Positive Control, Enzyme-linked Immunosorbent Assay, Negative Control, Expressing, Northern Blot

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 6. Induction of VSMC migration by resistin. VSMCs in serum-free media were incubated with 10¨100 ng/mL resistin for 24 h or with 10% FBS for 24 h as a positive control. Cell migration was detected by scratched wound assays and results are presented as fold changes versus the negative control. The data illustrated on the graph represent meansTSEM of three different experiments. *p <0.05 vs. without resistin treatment.

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Migration, Incubation, Positive Control, Negative Control

Journal: Cardiovascular research

Article Title: Resistin is secreted from macrophages in atheromas and promotes atherosclerosis.

doi: 10.1016/j.cardiores.2005.09.015

Figure Lengend Snippet: Fig. 5. Induction of ET-1 in HUVECs after resistin treatment. HUVECs were incubated with 10¨100 ng/mL of resistin for 4¨24 h or with 10 ng/mL of TNF-a for 12 h as a positive control. ET-1 mRNA levels were analyzed by Northern blotting and normalized against GAPDH mRNA, and are presented as fold changes versus the negative control. The data illustrated on the graph represent the meansTSEM of three different experiments. *p <0.05 vs. without resistin treatment.

Article Snippet: The sectionswere then incubated with primary antibodies, namely, monoclonal antihuman resistin (R&D Systems, Minneapolis, clone 184335), and anti-human CD68 (DakoCytomation, Glostrup, Denmark, clone KP1), the latter of which is a marker for monocytes/macrophages [22].

Techniques: Incubation, Positive Control, Northern Blot, Negative Control