rabbit anti mglur5 1  (R&D Systems)

Journal: Cancers

Article Title: Expression of mGluR5 in Pediatric Hodgkin and Non-Hodgkin lymphoma—A Comparative Analysis of Immunohistochemical and Clinical Findings Regarding the Association between Tumor and Paraneoplastic Neurological Disease

doi: 10.3390/cancers16132452

Figure Lengend Snippet: DAB staining of the images of HL ( a , b ), NHL ( c – f ) and control ( g – i ) tissues with mGluR5 antibody shows positive cells in the lymphoma sections and no specific staining in tumor free lymph nodes. ( a ) Examples of HL sections with defined positive tumor cells corresponding in morphology and localization to CD30+ H-RS cells, suggesting elevated expression of mGluR5 in H-RS cells. ( b ) HL sections without individually highlighted tumor cells but with a more homogenous staining pattern. ( c – f ) Examples of NHL with mGluR5+ cells and differing intensities between histological subtypes; ( c ) ALCL ( d ) DLBCL ( e ) Burkitt lymphoma ( f ) T-LBL. ( g – i ) Healthy lymph node tissues from three individual children do not show any positive staining for mGluR5 in lymphocytes. ( a – f ) Original objective lens magnification was ×20 for all lymphoma images; ×5 for ( g ) and ×10 for ( h , i ). The scale bar indicates 100 μm in all images.

Article Snippet: We performed staining with anti-mGluR5 primary antibody (1:200, Cat# PA5-33823 Invitrogen Waltham, MA, USA) on tissue sections from each patient.

Techniques: Staining, Control, Expressing

Journal: Cancers

Article Title: Expression of mGluR5 in Pediatric Hodgkin and Non-Hodgkin lymphoma—A Comparative Analysis of Immunohistochemical and Clinical Findings Regarding the Association between Tumor and Paraneoplastic Neurological Disease

doi: 10.3390/cancers16132452

Figure Lengend Snippet: Examples of immunofluorescence co-staining with anti-CD30 and -mGluR5 antibodies on HL and NHL tissues with corresponding DAB staining images show mGluR5 expression in CD30+ tumor cells and examples of autofluorescent signal. Numbers #1-#30 belong to HL and #31-#62 to NHL samples. All HL sections except #10 and #17 show clearly double stained H-RS cells. The pattern for mGluR5 in #10 and #17 is more diffuse and no (elevated) expression is seen in CD30+ cells, both sections showed a DAB staining pattern for mGluR5 depicted in b. #41 is an example of a CD30+ NHL (ALCL) with a high number of CD30+ cells and visible mGluR5 staining, which does not necessarily colocalize with CD30 in the IF images. Examples of autofluorescence are seen in #10 and #8 with multiple erythrocytes with excessive yellow staining due to exact overlap of red and green signal. The original objective lens magnification was 20× for all images, including DAB and IF stainings. The scale bar indicates 100 μm.

Article Snippet: We performed staining with anti-mGluR5 primary antibody (1:200, Cat# PA5-33823 Invitrogen Waltham, MA, USA) on tissue sections from each patient.

Techniques: Immunofluorescence, Staining, Expressing

Journal: Cancers

Article Title: Expression of mGluR5 in Pediatric Hodgkin and Non-Hodgkin lymphoma—A Comparative Analysis of Immunohistochemical and Clinical Findings Regarding the Association between Tumor and Paraneoplastic Neurological Disease

doi: 10.3390/cancers16132452

Figure Lengend Snippet: Image analysis of immunofluorescence co-staining: workflow visualization. The first row depicts the analysis of the total cell count by segmentation of the individual cell bodies using the DAPI stained nuclei. A standardized Gaussian blur filter was used to smooth out the intensities so that each nucleus could be automatically marked using the intensity maxima values. We then created a mask of the segmented nuclei, where each image pixel was assigned to a segment. We measured the created areas and counted each region < 500 µm 2 as an independent cell. Each cell segment was saved as a ROI and given a serial number. The next rows show the process of thresholding for the creation of binary masks of the CD30 and mGluR5 stainings by subtracting background staining and autofluorescence recorded identified in control channel images. We created an overlap of the CD30 and mGluR5 binary masks using the AND parameter to visualize the areas of co-staining. The masks were then merged, and positive cell-segments (=including black pixels) were automatically counted. During this process we accepted the false-positive counting of neighboring cell bodies in case of staining overlap due to the uniformly used method. The scale bar indicates 100 μm.

Article Snippet: We performed staining with anti-mGluR5 primary antibody (1:200, Cat# PA5-33823 Invitrogen Waltham, MA, USA) on tissue sections from each patient.

Techniques: Immunofluorescence, Staining, Cell Counting, Control

Journal: Cancers

Article Title: Expression of mGluR5 in Pediatric Hodgkin and Non-Hodgkin lymphoma—A Comparative Analysis of Immunohistochemical and Clinical Findings Regarding the Association between Tumor and Paraneoplastic Neurological Disease

doi: 10.3390/cancers16132452

Figure Lengend Snippet: Heterogeneous numbers of CD30+ cells in HL and NHL tissues and expression of mGluR5 in CD30+ tumor cells. The x-axis shows the serial number of the tissue samples (#), where samples #1–30 belong to HL cases (left half) and samples #31–62 belong to NHL cases (right half). In the upper panel are the box plots depicting the image analysis results for absolute cell counts per tissue sample, showing varying amounts of CD30+ tumor cells independent from the total cell count. The box plots in the lower panel in light red depict the results in percent for CD30+ cells relative to the total cell count. In light yellow, the percentage of double positive (mGluR5 and CD30) tumor cells is shown relative to the total amount of CD30+ cells. For the NHL cases, the results are shown only for tissues with actual CD30-expression marked by an asterisk (*). Each box plot contains the results of n = 3 independent measurements of ×20 magnification images and the median is depicted as the center line.

Article Snippet: We performed staining with anti-mGluR5 primary antibody (1:200, Cat# PA5-33823 Invitrogen Waltham, MA, USA) on tissue sections from each patient.

Techniques: Expressing, Cell Counting

Journal: Cancers

Article Title: Expression of mGluR5 in Pediatric Hodgkin and Non-Hodgkin lymphoma—A Comparative Analysis of Immunohistochemical and Clinical Findings Regarding the Association between Tumor and Paraneoplastic Neurological Disease

doi: 10.3390/cancers16132452

Figure Lengend Snippet: Sorting of image analysis results shows no correlation between CD30 and mGluR5 expression (in co-expressing lymphoma cells). We suggest the division into two groups of low and high mGluR5 expression in CD30+ tumor cells. The left panels of the graph show the mean results per tissue for the percentage of CD30+ cells and are sorted from highest to lowest. The right panels are sorted by the percentage of mGluR5 expression. All column plots show the mean of the measured parameter, the box plots include the individual measurements per tissue and indicate the minimum, maximum and median, as well as the 25th and 75th percentiles. The right panels include the 40% cut-off (red line) for dividing the subgroups. An inset box plot indicates a significant difference between the means of the two groups. Considering the highly sensitive image analysis algorithm, results of below 40% can be dismissed as low expression of the antigen. This corresponds with the DAB staining results.

Article Snippet: We performed staining with anti-mGluR5 primary antibody (1:200, Cat# PA5-33823 Invitrogen Waltham, MA, USA) on tissue sections from each patient.

Techniques: Expressing, Staining

Journal: PLoS ONE

Article Title: Transplacental Exposure to AZT Induces Adverse Neurochemical and Behavioral Effects in a Mouse Model: Protection by L-Acetylcarnitine

doi: 10.1371/journal.pone.0055753

Figure Lengend Snippet: ( A ) Representatives of immunoblots with mGlu1a and mGlu5 receptor antibodies showed a 142 kDa and 130 kDa bands, corresponding to receptor monomers, respectively. Blots with antibodies recognize an epitope common to mGlu2 receptors monomer(s) (100 kDa) and mGlu3 receptors dimers (206 kDa). ( B ) Results are expressed as the ratio of the optical density (OD) of the mGluR1a, mGluR5 or mGlu2/3 band and the β-actin band. The AZT prenatal treatment caused a significant reduction of mGlu1a and mGlu5 receptors expression in respect to Control mice (* p<0.05, Student's t-test) counteracted by LAC treatment as AZT+LAC mice differ from Control. No differences between groups were found in the mGluR2/3 (summary of OD monomers and dimers). Values are expressed as means ± S.E.M. (n = 6 mice per group).

Article Snippet: Subsequently, blots were incubated overnight with rabbit anti-mGluR1a (1∶1000), anti-mGluR5 (1∶1000), anti-mGluR2/3 (1∶1000), anti-NR1 (1∶1000), mouse antiGlu1 (1∶500) (Upstate Biotechnology) and mouse antiGlu2 (1∶500; Chemicon International) in blocking solution at 4°C.

Techniques: Western Blot, Expressing