anti rat igg whole molecule  (Millipore)

Journal: Cardiovascular Diabetology

Article Title: Immunization with advanced glycation end products modified low density lipoprotein inhibits atherosclerosis progression in diabetic apoE and LDLR null mice

doi: 10.1186/s12933-014-0151-6

Figure Lengend Snippet: Serum titers of IgG, IgM, IgG1 and IgG2a antibodies. A . ELISA analysis of titers of IgG, IgM, and specific anti-AGE-LDL IgG and IgM antibodies in sera from diabetic apoE−/− and LDLR−/− mice (n = 10 per group) treated with Alum adjuvant (bar with stripes), AGE-LDL + Alum adjuvant (black bar) or PBS controls (white bar). B . ELISA analysis of specific IgG1 and IgG2a antibodies against AGE-LDL in sera from diabetic apoE−/− and LDLR−/− mice treated with Alum adjuvant, AGE-LDL + Alum adjuvant or PBS controls. C shows the ratio of serum IgG1/IgG2a antibodies among three groups. *indicates p

Article Snippet: After washing, bound antibodies were detected by using biotinylated goat anti-mouse IgM (AP500B, Millipore, USA) or IgG (B7264, Sigma, USA) or rat anti-mouse IgG1 or IgG2a secondary antibodies (RMG115 and RMG2a15, Millipore, USA) that were incubated for 2 hours at room temperature.

Techniques: Enzyme-linked Immunosorbent Assay, Mouse Assay

Journal: Scientific Reports

Article Title: N-Myc-Interacting Protein Negatively Regulates TNF-α-Induced NF-κB Transcriptional Activity by Sequestering NF-κB/p65 in the Cytoplasm

doi: 10.1038/s41598-017-15074-5

Figure Lengend Snippet: NMI inhibits the nuclear translocation of NF-κB/p65 after TNF-α stimulation. (A) HeLa cells were transfected with control or Myc-NMI plasmids, and at 24 h posttransfection, the cells were left untreated or treated with TNF-α (10 ng/ml) for 60 min. The cells were fixed and incubated with anti-p65 or anti-Myc Ab and then stained with rhodamine-conjugated anti-mouse IgG (red) or FITC-conjugated anti-rabbit Ab (green). The same slide was also stained with DAPI to show the nucleus. The expression and localization of p65 and Myc-NMI was determined by confocal immunofluorescence analysis. The percentage of the cells expressing p65 in the nucleus among the cells that express Myc-NMI and the cells that don’t express Myc-NMI was calculated in the bottom panel. (B) HeLa cells were transfected with control or Myc-NMI plasmids, and at 24 h posttransfection, the cells were left untreated or treated with TNF-α (10 ng/ml) for 60 min. The cells were then harvested and fractionated into the nuclear and cytoplasmic fractions, and the fractions were immunoblotted with anti-NMI and anti-p65 Abs. The quantification by densitometry of western blots of p65 bands without or with Myc-NMI in TNF-α-stimulated nuclear fractions was shown in the right panel. The results are representative of three independent experiments, and the error bars represent the SD.**p

Article Snippet: Immunocomplexes were resolved by SDS-PAGE, and Western blotting was performed with the following Abs: anti-FLAG M2 and anti-actin monoclonal Abs (Sigma-Aldrich); anti-HA (Y-11), anti-IκB (C-20) and anti-NMI (N-16) polyclonal Abs, anti-Myc (9E10), anti-GFP (B-2) mAb (Santa Cruz Biotechnology); NF-κB/p65 (L8F6) mAb (Cell signaling); horseradish peroxidade-conjugated goat anti-mouse, goat anti-rabbit and donkey-anti goat (Thermo Scientific, Waltham, MA), and FITC-conjugated anti-rabbit IgG or rhodamine-conjugated anti-mouse IgG (Millipore).

Techniques: Translocation Assay, Transfection, Incubation, Staining, Expressing, Immunofluorescence, Western Blot

Journal: PLoS ONE

Article Title: Middle Infrared Radiation Induces G2/M Cell Cycle Arrest in A549 Lung Cancer Cells

doi: 10.1371/journal.pone.0054117

Figure Lengend Snippet: Effect of MIR exposure on DNA double strain breaks in A549 cells. Cells were seeded onto the glass coverslip in 12-well plate, exposure by MIR for 48 hours in the presence or absence of 10 mM N-Acetyl-Cysteine (NAC). Cells were treated with NAC for 1 h prior to MIR exposure (NAC 1 h) or cotreated throughout the exposure for 48 h (NAC). Cells were fixed for staining and visualized by fluorescence microscopy. 53BP1 was labeled with rabbit anti-53BP1 antibody and corresponded FITC–conjugated anti-rabbit IgG antibody (green), γ-H2AX was labeled with mouse anti-γ-H2AX antibody following corresponded PE–conjugated anti-mouse IgG antibody (red), and nuclei were labeled with DAPI (blue). Scale bar represents 10 µm.

Article Snippet: After washed with PBST (PBS containing 0.05% Tween-20, Sigma-Aldrich, St Louis, MO, USA) three times, cells were labeled with corresponding secondary anti-mouse IgG-Alexa 488 (Invitrogen, Carlsbad, CA; 1∶1000), anti-mouse IgG-PE (Abcam; 1∶1000), or secondary anti-rabbit FITC-IgG (Millipore; 1∶200) for 1 hour in the dark at room temperature.

Techniques: Staining, Fluorescence, Microscopy, Labeling

Journal: PLoS ONE

Article Title: Middle Infrared Radiation Induces G2/M Cell Cycle Arrest in A549 Lung Cancer Cells

doi: 10.1371/journal.pone.0054117

Figure Lengend Snippet: Effect of MIR exposure on the actin filaments and focal adhesions of A549 cells. Cells were seeded onto glass coverslips in 12-well plates, exposed to MIR for 48 hours, fixed for staining and visualized by fluorescence microscopy. Actin filaments were tagged with rhodamine-labeled phalloidin (red), vinculin was labeled with mouse anti-vinculin antibody and the corresponding FITC– conjugated secondary anti-mouse IgG antibody (green), and nuclei were stained with DAPI (blue). Scale bar represents 10 µm. Arrows indicate the position of vinculin.

Article Snippet: After washed with PBST (PBS containing 0.05% Tween-20, Sigma-Aldrich, St Louis, MO, USA) three times, cells were labeled with corresponding secondary anti-mouse IgG-Alexa 488 (Invitrogen, Carlsbad, CA; 1∶1000), anti-mouse IgG-PE (Abcam; 1∶1000), or secondary anti-rabbit FITC-IgG (Millipore; 1∶200) for 1 hour in the dark at room temperature.

Techniques: Staining, Fluorescence, Microscopy, Labeling

Journal: Parasites & Vectors

Article Title: Immunolocalization of TSOL18 and TSOL45-1A, the successful protective peptides against porcine cysticercosis, in Taenia solium oncospheres

doi: 10.1186/1756-3305-4-3

Figure Lengend Snippet: Taenia solium oncospheres obtained from a tapeworm carrier are seen under contrast phase microscopy, PI, FITC and a merged image of both stains . The first row corresponds to oncospheres incubated with anti GST IgG as a negative control; rows 2 and 3 include images obtained with the specific anti TSOL18 and anti TSOL45-1A antibodies, respectively. The arrows in the first column point to the hooks of the oncospheres that was used for focus. Positive reactions were detected with anti-TSOL18 and anti-TSOL45-1A antibodies only on the surface of the oncospheres, as seen in the merged image. The high intensity emission of PI in FITC sections caused non-specific background fluorescence on the same internal structures seen in PI sections.

Article Snippet: Whole immunoglobulin G (IgG) was purified with a commercial kit (Montage Antibody Purification Prosep-A, Millipore, Bedford, MA, USA), the purified IgG was adjusted at 1 mg/ml.

Techniques: Microscopy, Incubation, Negative Control, Fluorescence