rabbit anti igg3  (Abcam)

 
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    Name:
    Rabbit F ab 2 Anti Rat IgG H L TR
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    Catalog Number:
    ab6247
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    Structured Review

    Abcam rabbit anti igg3
    <t>IgG3–IgM</t> co-localization on IgG3+IgM+ B cells. a , Imaging cytometry (top right) of CD20+ B cells from a chronically infected HIV-viremic individual, gated (left) on populations with different levels of IgG3 and IgM (arrows at left); below, frequency of IgG3–IgM co-localization, based on quantitative analysis of the relative intensity of two fluorescent images. b , Pearson’s co-localization coefficient ( R ) for various protein pairs (horizontal axis), as measured by TIRF microscopy of B cells from a chronically infected HIV-viremic individual with high-intensity IgG3 on IgM + B cells (H-) and one with low-intensity IgG3 on IgM + B cells (L-), stained for tIgG (with polyclonal anti-IgG), IgG3, IgM and CD45; R values were calculated pairwise for each marker and for each source of cells. c , R values for IgG3–IgM co-localization on the TLM B cells and naive B cells (horizontal axis) of a chronically infected HIV-viremic individual with high-intensity IgG3 on IgM + B cells. Results in b , c are presented as ‘box and whiskers’ plots (middle line, mean; box limits, inter-quartile distance; extended lines, Tukey’s method); in parentheses (above horizontal axis), number of cells imaged for each analysis. **** P

    https://www.bioz.com/result/rabbit anti igg3/product/Abcam
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti igg3 - by Bioz Stars, 2020-09
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    Images

    1) Product Images from "IgG3 regulates tissue-like memory B cells in HIV-infected individuals"

    Article Title: IgG3 regulates tissue-like memory B cells in HIV-infected individuals

    Journal: Nature immunology

    doi: 10.1038/s41590-018-0180-5

    IgG3–IgM co-localization on IgG3+IgM+ B cells. a , Imaging cytometry (top right) of CD20+ B cells from a chronically infected HIV-viremic individual, gated (left) on populations with different levels of IgG3 and IgM (arrows at left); below, frequency of IgG3–IgM co-localization, based on quantitative analysis of the relative intensity of two fluorescent images. b , Pearson’s co-localization coefficient ( R ) for various protein pairs (horizontal axis), as measured by TIRF microscopy of B cells from a chronically infected HIV-viremic individual with high-intensity IgG3 on IgM + B cells (H-) and one with low-intensity IgG3 on IgM + B cells (L-), stained for tIgG (with polyclonal anti-IgG), IgG3, IgM and CD45; R values were calculated pairwise for each marker and for each source of cells. c , R values for IgG3–IgM co-localization on the TLM B cells and naive B cells (horizontal axis) of a chronically infected HIV-viremic individual with high-intensity IgG3 on IgM + B cells. Results in b , c are presented as ‘box and whiskers’ plots (middle line, mean; box limits, inter-quartile distance; extended lines, Tukey’s method); in parentheses (above horizontal axis), number of cells imaged for each analysis. **** P
    Figure Legend Snippet: IgG3–IgM co-localization on IgG3+IgM+ B cells. a , Imaging cytometry (top right) of CD20+ B cells from a chronically infected HIV-viremic individual, gated (left) on populations with different levels of IgG3 and IgM (arrows at left); below, frequency of IgG3–IgM co-localization, based on quantitative analysis of the relative intensity of two fluorescent images. b , Pearson’s co-localization coefficient ( R ) for various protein pairs (horizontal axis), as measured by TIRF microscopy of B cells from a chronically infected HIV-viremic individual with high-intensity IgG3 on IgM + B cells (H-) and one with low-intensity IgG3 on IgM + B cells (L-), stained for tIgG (with polyclonal anti-IgG), IgG3, IgM and CD45; R values were calculated pairwise for each marker and for each source of cells. c , R values for IgG3–IgM co-localization on the TLM B cells and naive B cells (horizontal axis) of a chronically infected HIV-viremic individual with high-intensity IgG3 on IgM + B cells. Results in b , c are presented as ‘box and whiskers’ plots (middle line, mean; box limits, inter-quartile distance; extended lines, Tukey’s method); in parentheses (above horizontal axis), number of cells imaged for each analysis. **** P

    Techniques Used: Imaging, Cytometry, Infection, Microscopy, Staining, Marker

    IgG3+IgM+ B cells have reduced responses to BCR stimulation. a , Flow cytometry (top) of CD20-gated B cells of a chronically infected HIV-viremic individual, showing expression of CD27 and CD21 (far left) and the IgG3 and IgD profiles of the corresponding TLM B cell and naive B cell populations (middle and right); below, calcium uptake by IgG3 − IgD + and IgG3 + IgD + (far left margin) TLM B cells (left) and naive B cells (right) before (Unstimulated) and after (Anti-IgM) 2 min of incubation with F(ab’) 2 anti-IgM; the peak (number in plots) is the ratio of bound fluorescent Ca2 + indicator dye Indo-1 to unbound Indo-1. Data are representative of nine experiments. b , Calcium uptake by IgG3 + IgD + TLM B cells (TLM G3+D+), IgG3 − IgD + TLM B cells (TLM G3 − D + ), IgG3 + IgD + naive B cells (N G3+D+) and IgG3 − IgD + naive B cells (N G3 − D + ) from chronically infected HIV-viremic individuals with moderate- to high-intensity IgG3 on IgM + B cells, presented as the peak ratio (as in a ) after incubation with anti-IgM relative to that ratio in unstimulated cells (‘fold’ values). c , d , Quantification of phosphorylated (p-) Syk, PLC-γ2 and Btk (measured by flow cytometry) in B cell populations as in b (horizontal axis) after 2 min of incubation with F(ab’) 2 anti-IgM ( c ) or in the absence of such incubation with anti-IgM ( d ); results in c after incubation with anti-IgM are presented relative to baseline (‘fold’ values). Each symbol ( b – d ) represents an individual subject ( n = 9 ( b ) or n = 11 ( c , d )); small horizontal lines indicate the median. * P
    Figure Legend Snippet: IgG3+IgM+ B cells have reduced responses to BCR stimulation. a , Flow cytometry (top) of CD20-gated B cells of a chronically infected HIV-viremic individual, showing expression of CD27 and CD21 (far left) and the IgG3 and IgD profiles of the corresponding TLM B cell and naive B cell populations (middle and right); below, calcium uptake by IgG3 − IgD + and IgG3 + IgD + (far left margin) TLM B cells (left) and naive B cells (right) before (Unstimulated) and after (Anti-IgM) 2 min of incubation with F(ab’) 2 anti-IgM; the peak (number in plots) is the ratio of bound fluorescent Ca2 + indicator dye Indo-1 to unbound Indo-1. Data are representative of nine experiments. b , Calcium uptake by IgG3 + IgD + TLM B cells (TLM G3+D+), IgG3 − IgD + TLM B cells (TLM G3 − D + ), IgG3 + IgD + naive B cells (N G3+D+) and IgG3 − IgD + naive B cells (N G3 − D + ) from chronically infected HIV-viremic individuals with moderate- to high-intensity IgG3 on IgM + B cells, presented as the peak ratio (as in a ) after incubation with anti-IgM relative to that ratio in unstimulated cells (‘fold’ values). c , d , Quantification of phosphorylated (p-) Syk, PLC-γ2 and Btk (measured by flow cytometry) in B cell populations as in b (horizontal axis) after 2 min of incubation with F(ab’) 2 anti-IgM ( c ) or in the absence of such incubation with anti-IgM ( d ); results in c after incubation with anti-IgM are presented relative to baseline (‘fold’ values). Each symbol ( b – d ) represents an individual subject ( n = 9 ( b ) or n = 11 ( c , d )); small horizontal lines indicate the median. * P

    Techniques Used: Flow Cytometry, Cytometry, Infection, Expressing, Incubation, Planar Chromatography

    Soluble IgG3 binds to IgM-expressing B cells of HIV-viremic individuals in vivo. a,b, Flow cytometry of B cells isolated from the peripheral blood of HIV-negative, HIV-aviremic and HIV-viremic individuals (above plots), CD20 gated (to exclude plasmablasts, which typically express CD19 but not CD20), and stained for IgG3 and tIgG ( a ) or IgM ( b ). Numbers in quadrants indicate percent cells in each throughout. c , Flow cytometry showing immunoglobulin light-chain expression (middle and right) on IgG3 + IgM + and combined single immunoglobulin-positive (Ig + ) B cells of an HIV-viremic individual. d , Flow cytometry showing staining for tIgG, IgG1, IgG3 and IgM of B cells obtained from an HIV-viremic individual and treated with trypsin (bottom row) or not (top row) for 5 min at 37°C. e , Expression of immunoglobulin-encoding mRNA (key) by B cells sorted by the surface expression as tIgG + IgM − , tIgG − IgM + or IgG3 + IgM + (horizontal axis); band intensity was normalized to that of control mRNA encoding β -actin. Data are representative of 83 experiments (HIV-negative), 17 experiments (HIV-aviremic) or 75 experiments (HIV-viremic) ( a,b ), 17 ( c ) or 13 ( d ) experiments, or three independent experiments ( e ).
    Figure Legend Snippet: Soluble IgG3 binds to IgM-expressing B cells of HIV-viremic individuals in vivo. a,b, Flow cytometry of B cells isolated from the peripheral blood of HIV-negative, HIV-aviremic and HIV-viremic individuals (above plots), CD20 gated (to exclude plasmablasts, which typically express CD19 but not CD20), and stained for IgG3 and tIgG ( a ) or IgM ( b ). Numbers in quadrants indicate percent cells in each throughout. c , Flow cytometry showing immunoglobulin light-chain expression (middle and right) on IgG3 + IgM + and combined single immunoglobulin-positive (Ig + ) B cells of an HIV-viremic individual. d , Flow cytometry showing staining for tIgG, IgG1, IgG3 and IgM of B cells obtained from an HIV-viremic individual and treated with trypsin (bottom row) or not (top row) for 5 min at 37°C. e , Expression of immunoglobulin-encoding mRNA (key) by B cells sorted by the surface expression as tIgG + IgM − , tIgG − IgM + or IgG3 + IgM + (horizontal axis); band intensity was normalized to that of control mRNA encoding β -actin. Data are representative of 83 experiments (HIV-negative), 17 experiments (HIV-aviremic) or 75 experiments (HIV-viremic) ( a,b ), 17 ( c ) or 13 ( d ) experiments, or three independent experiments ( e ).

    Techniques Used: Expressing, In Vivo, Flow Cytometry, Cytometry, Isolation, Staining

    Related Articles

    Western Blot:

    Article Title: p73 is required for endothelial cell differentiation, migration and the formation of vascular networks regulating VEGF and TGFβ signaling
    Article Snippet: .. Western blotting was performed as previously described with the following primary antibodies: rabbit anti-pSmad1/5 (Ser463/465) 1 : 1000 (Cell Signaling, Danvers, MA, USA), rabbit anti Smad1 1 : 1000 (Cell Signaling), mouse anti-pERK 1 : 1000 (Santa Cruz Biotechnology), rabbit anti-ERK1 : 10 000 (Santa Cruz Biotechnology), rabbit anti-VEGF-A 1 : 1000 (Abcam) mouse anti DNp73 1 : 1000 (Imgenex, San Diego, CO, USA), and rabbit anti Nanog 1 : 1000 (Chemicon, Billerica, TX, USA) followed by the appropriate HRP-conjugated secondary antibodies (Pierce, Waltham, MA, USA). .. The enhanced chemiluminescence was detected with Super Signal West-Pico Chemiluminiscent Substrate (Pierce).

    Incubation:

    Article Title: IgG3 regulates tissue-like memory B cells in HIV-infected individuals
    Article Snippet: .. Membranes were sequentially incubated with goat anti-IgM-HRP (Southern Biotech; Cat# 2020–05) or primary antibody rabbit anti-C1q (Novus; Cat# H00000712-D01P), mouse anti-CRP (R & D Systems; clone 232026), rabbit anti-CD32 (GeneTex; Cat# GTX133371) or rabbit anti- IgG3 (Abcam; clone EPR4419), followed by the secondary antibody, goat anti-rabbit-HRP (Bio-Rad; Cat# 1662408EDU) or goat anti-mouse-HRP (Bio-Rad; Cat# 1000450), diluted in 4% milk powder in 0.1% Tween in PBS. .. B cells (5 × 105 ) of HIV-negative individuals were incubated for 20 min at 37 °C with 5 μ g of heat-aggregated IgG3 (Sigma; Cat# I5654) or (Bio-Rad; clone ), PEG-IgG3, fractions SEC-1 and SEC-2 or agarose-bead-enriched IgG3, isolated from the serum of HIV-viremic individuals.

    Article Title: Homeostatic Control of Sebaceous Glands by Innate Lymphoid Cells Regulates Commensal Bacteria Equilibrium
    Article Snippet: .. Epidermal sheets were then incubated with goat anti-LRIG1 (polyclonal, R & D) at 1:20, rabbit anti-MGST1 (EPR7934, Abcam) at 1:200, and rat anti-Ki-67 (11F6, Biolegend) antibodies at 1:100 overnight at 4°C. .. The samples were washed three times with PBS, 5 min each, then incubated with Alexa Fluor 488-conjugated anti-goat (Thermo Fisher Scientific), and Alexa Fluor 568-conjugated anti-rabbit (Thermo Fisher Scientific) antibody for 45 min at room temperature.

    Article Title: Phospho-Tau Accumulation and Structural Alterations of the Golgi Apparatus of Cortical Pyramidal Neurons in the P301S Tauopathy Mouse Model
    Article Snippet: .. The sections were then incubated for 48 h at 4°C in the same stock solution containing the following primary antibodies in the combinations indicated: rabbit anti-MG160 (Abcam, 1 : 100), rabbit anti-Grasp65 (Abcam, 1 : 500), mouse anti human tau (Abcam, T13, 1 : 5000), rabbit anti-NeuN (Millipore, 1 : 2000) and mouse anti-phospho-PHF-tau pSer202+ Thr205 antibody (AT8, 1 : 2000, Pierce Endogen). .. After rinsing in PB, the sections were first incubated for 2 h at room temperature in biotinylated goat anti-rabbit antibody (1 : 200) to amplify the GA immunoreactivity signal.

    Article Title: Combination therapy with F5/35 fiber chimeric conditionally replicative adenoviruses expressing IL‐24 enhances the antitumor effect of temozolomide against melanoma, et al. Combination therapy with F5/35 fiber chimeric conditionally replicative adenoviruses expressing IL‐24 enhances the antitumor effect of temozolomide against melanoma
    Article Snippet: .. Proteins were transferred on to a nitrocellulose membrane and incubated at 4°C overnight, together with the following primary antibodies: rabbit anti‐CAR, rabbit anti‐CD46 (dilution 1:500; Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit anti‐IL‐24 (dilution 1:500; Bioss, Beijing, China); mouse anti‐Bcl‐2, rabbit anti‐Bax, rabbit anti‐Pro‐caspase‐3, and mouse anti‐p53 (dilution 1:1000; Absci, College Park, MD, USA); rabbit anti‐MGMT (dilution 1:1000; Abcam, Cambridge, MA, USA); mouse anti‐E1A (dilution 1:500, EMD Millipore, Burlington, MA, USA); or rabbit anti‐β‐actin (dilution 1:5000; Bioworld, St Louis Park, MN, USA). .. The membranes were then washed and incubated for 2 hours with alkaline phosphatase‐conjugated secondary antibodies (goat anti‐rabbit, dilution 1:1000, Zhongshan, Beijing, China; goat anti‐mouse, dilution 1:1000, Zhongshan) in Tris‐buffered saline with Tween‐20 (TBST) and developed using a nitro‐blue tetrazolium and 5‐Bromo‐4‐chloro‐3' prime symbol‐indolyl phosphate (NBT/BCIP) color development substrate (Promega, Madison, WI, USA).

    Article Title: Glycyrrhizin Protects Mice Against Experimental Autoimmune Encephalomyelitis by Inhibiting High-Mobility Group Box 1 (HMGB1) Expression and Neuronal HMGB1 Release
    Article Snippet: .. The membranes were blocked with 5% non-fat dry milk in TBS solution for 1 h at room temperature, followed by incubation overnight with primary antibodies, including rabbit anti-HMGB1 (1:8,000; Abcam), mouse anti-β-actin (1:500; Santa Cruz Biotechnology, Dallas, TX, USA), and rabbit anti-Lamin B1 (1:500; Proteintech Group, Inc., Chicago, IL, USA) antibodies. .. The membrane was then incubated with a horseradish peroxidase-conjugated anti-IgG (1:2,000; CWBIO) for 1 h at room temperature.

    Article Title: Morphometric alterations of Golgi apparatus in Alzheimer's disease are related to tau hyperphosphorylation
    Article Snippet: .. The sections were then incubated for 48 h at 4 °C in the same stock solution containing the following primary antibodies in the combinations indicated: rabbit anti-MG160 (Abcam, 1:100), mouse anti-NeuN (Chemicon, 1:2000), mouse phospho-PHF-tau pSer202 + Thr205 antibody (AT8, 1:2000, Pierce Endogen). .. After rinsing in PB, the sections were incubated for 2 h at room temperature in the appropriate combinations of Alexa 488- or Alexa 594-conjugated goat anti-mouse or goat anti-rabbit antibodies (1:2000; Molecular Probes, Eugene, OR, USA).

    Immunocytochemistry:

    Article Title: Dynein light chain regulates axonal trafficking and synaptic levels of Bassoon
    Article Snippet: .. Antibodies and stains The following antibodies were used: rabbit antibodies against DLC (DLC8; diluted 1:40,000 for WBs and 1:1,000 for immunocytochemistry [ICC]; Abcam), DLC1/2 (diluted 1:1,000 for ICC; gift from M. Sheng, Massachusetts Institute of Technology, Cambridge, MA; ), synaptophysin (diluted 1:5,000 for ICC; gift from R. Jahn, Max Plank Institute for Biophysical Chemistry, Göttingen, Germany), Bassoon (sap7f; diluted 1:2,000 for ICC; ), GFP (diluted 1:10,000 for WBs; Abcam), myosin V (diluted 1:1,000 for ICC; Sigma-Aldrich), Munc13-1 (diluted 1:1,000 for ICC; Synaptic Systems GmbH), and synapsin (diluted 1:2,000 for ICC; Synaptic Systems GmbH). .. Mouse antibodies used were against cytochrome c (diluted 1:100 for ICC; BD), IC74 (diluted 1:2,000 for WBs and 1:50 for ICC; Millipore), Flag M2 (diluted 1:10,000 for WBs; Sigma-Aldrich), kinesin heavy chain (H2; diluted 1:2,000 for WBs; Millipore), c-myc (9E10; diluted 1:1,000 for WBs; Santa Cruz Biotechnology, Inc.), α-tubulin (diluted 1:1,000; Sigma Aldrich), synapsin (diluted 1:2,000 for ICC; Synaptic Systems GmbH), PSD-95 (diluted 1:1,000–2,000 for ICC; Millipore), and Bassoon (mab7f; diluted 1:2,000 for WBs and 1:1,000 for ICC; Assay Designs).

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  • 91
    Abcam anti asialo gm1
    Domain formation in B. burgdorferi as determined by immunogold TEM analysis requires raft-supporting sterols. A. Representative negative-stain TEM images of B. burgdorferi substituted with the indicated sterols and probed for sterol glycolipids using a rabbit antibody to <t>asialo-GM1</t> followed by a secondary anti-rabbit antibody conjugated to 6 nm colloidal gold. Micrographs show electron dense regions, which are portions of B. burgdorferi and show associated gold particles. Two images (of about 400 nm long segments) are shown for each sterol. Top row, sterols strongly supporting ordered domain formation; middle row, sterols with an intermediate ability to form ordered domains; bottom row, sterols that inhibit ordered domain formation [22] , [26] – [28] . Boxes highlight sterol glycolipid clusters. Bars = 100 nm. TEM micrographs for additional sterols are shown in Fig. S3 in Text S1 . B. Pooled “K-function” analysis of TEM experiments. The spatial distribution of gold particles is presented as curves representing the mean values of L(r)-r from images of three different bacteria from three independent sterol substitution experiments. Values of L(r)-r above the CI (95% confidence level, dashed line) indicate clustering (i.e. domain formation) of the sterol glycolipid at that specific length scale.
    Anti Asialo Gm1, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti asialo gm1/product/Abcam
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti asialo gm1 - by Bioz Stars, 2020-09
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    89
    Abcam anti lat 1 mab
    Microscopy images (×40) of immunofluorescent staining of <t>LAT-1</t> (tumour (a) , granuloma (d) ), GLUT-1 (tumour (b) , granuloma (e) ) and H E staining (tumour (c) , granuloma (f) ). White arrowhead : cancer cell; black arrow : lymphocyte infiltration; white arrow : epithelioid cell granuloma. Bars indicate 50 μm.
    Anti Lat 1 Mab, supplied by Abcam, used in various techniques. Bioz Stars score: 89/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti lat 1 mab/product/Abcam
    Average 89 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    anti lat 1 mab - by Bioz Stars, 2020-09
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    99
    Abcam anti rabbit igg
    Recombinant ACKR3 and <t>AVPR1A</t> form heteromeric complexes. ( a ) Typical PLA images for the detection of individual receptors and receptor–receptor interactions in HEK293T cells transfected with DNA encoding HA- or FLAG-tagged receptors. Ctrl: cells transfected with pcDNA. Images show merged PLA/DAPI signals acquired from z -stack images ( n = 10; thickness 1 µm, bottom to top). Scale bars, 10 µm. ( b ) Quantification of PLA signals per cell as in ( a ). n = 3 independent experiments with n = 10 images per condition and experiment. ( c ) HEK293T cells expressing HA-AVPR1A and FLAG-ACKR3 were lysed (input), the lysate was immunoprecipitated (IP) with anti-HA, followed by immunoblotting (IB) to detect HA-AVPR1A (left) and FLAG-ACKR3 (right) in the IP samples. IP control: precipitate after incubation of cell lysates with <t>IgG-coupled</t> resin. ( d , e ) Intermolecular BRET assays. Cells were co-transfected with AVPR1A-hRluc plus EYFP (open circles) or ACKR3-EYFP (grey squares) at various acceptor : donor ratios ( d ) and with increasing amounts of AVPR1A-hRluc and ACKR3-EYFP at a constant ratio of 1 : 10 ( e ).
    Anti Rabbit Igg, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Domain formation in B. burgdorferi as determined by immunogold TEM analysis requires raft-supporting sterols. A. Representative negative-stain TEM images of B. burgdorferi substituted with the indicated sterols and probed for sterol glycolipids using a rabbit antibody to asialo-GM1 followed by a secondary anti-rabbit antibody conjugated to 6 nm colloidal gold. Micrographs show electron dense regions, which are portions of B. burgdorferi and show associated gold particles. Two images (of about 400 nm long segments) are shown for each sterol. Top row, sterols strongly supporting ordered domain formation; middle row, sterols with an intermediate ability to form ordered domains; bottom row, sterols that inhibit ordered domain formation [22] , [26] – [28] . Boxes highlight sterol glycolipid clusters. Bars = 100 nm. TEM micrographs for additional sterols are shown in Fig. S3 in Text S1 . B. Pooled “K-function” analysis of TEM experiments. The spatial distribution of gold particles is presented as curves representing the mean values of L(r)-r from images of three different bacteria from three independent sterol substitution experiments. Values of L(r)-r above the CI (95% confidence level, dashed line) indicate clustering (i.e. domain formation) of the sterol glycolipid at that specific length scale.

    Journal: PLoS Pathogens

    Article Title: Proving Lipid Rafts Exist: Membrane Domains in the Prokaryote Borrelia burgdorferi Have the Same Properties as Eukaryotic Lipid Rafts

    doi: 10.1371/journal.ppat.1003353

    Figure Lengend Snippet: Domain formation in B. burgdorferi as determined by immunogold TEM analysis requires raft-supporting sterols. A. Representative negative-stain TEM images of B. burgdorferi substituted with the indicated sterols and probed for sterol glycolipids using a rabbit antibody to asialo-GM1 followed by a secondary anti-rabbit antibody conjugated to 6 nm colloidal gold. Micrographs show electron dense regions, which are portions of B. burgdorferi and show associated gold particles. Two images (of about 400 nm long segments) are shown for each sterol. Top row, sterols strongly supporting ordered domain formation; middle row, sterols with an intermediate ability to form ordered domains; bottom row, sterols that inhibit ordered domain formation [22] , [26] – [28] . Boxes highlight sterol glycolipid clusters. Bars = 100 nm. TEM micrographs for additional sterols are shown in Fig. S3 in Text S1 . B. Pooled “K-function” analysis of TEM experiments. The spatial distribution of gold particles is presented as curves representing the mean values of L(r)-r from images of three different bacteria from three independent sterol substitution experiments. Values of L(r)-r above the CI (95% confidence level, dashed line) indicate clustering (i.e. domain formation) of the sterol glycolipid at that specific length scale.

    Article Snippet: For slot blots, fractions were loaded onto nitrocellulose and probed with anti-asialo GM1 (for sterol glycolipids, rabbit polyclonal IgG, AbCam) or monoclonal antibodies CB2 (anti-OspB, mouse IgG) or CB10 (anti-OspA, mouse IgG) , .

    Techniques: Transmission Electron Microscopy, Staining

    Microscopy images (×40) of immunofluorescent staining of LAT-1 (tumour (a) , granuloma (d) ), GLUT-1 (tumour (b) , granuloma (e) ) and H E staining (tumour (c) , granuloma (f) ). White arrowhead : cancer cell; black arrow : lymphocyte infiltration; white arrow : epithelioid cell granuloma. Bars indicate 50 μm.

    Journal: EJNMMI Research

    Article Title: Differentiation of malignant tumours from granulomas by using dynamic [18F]-fluoro-L-α-methyltyrosine positron emission tomography

    doi: 10.1186/s13550-015-0109-z

    Figure Lengend Snippet: Microscopy images (×40) of immunofluorescent staining of LAT-1 (tumour (a) , granuloma (d) ), GLUT-1 (tumour (b) , granuloma (e) ) and H E staining (tumour (c) , granuloma (f) ). White arrowhead : cancer cell; black arrow : lymphocyte infiltration; white arrow : epithelioid cell granuloma. Bars indicate 50 μm.

    Article Snippet: LAT-1 was stained by using an anti-LAT-1 mAb (rabbit IgG, synthetic peptide, Abcam).

    Techniques: Microscopy, Staining

    Recombinant ACKR3 and AVPR1A form heteromeric complexes. ( a ) Typical PLA images for the detection of individual receptors and receptor–receptor interactions in HEK293T cells transfected with DNA encoding HA- or FLAG-tagged receptors. Ctrl: cells transfected with pcDNA. Images show merged PLA/DAPI signals acquired from z -stack images ( n = 10; thickness 1 µm, bottom to top). Scale bars, 10 µm. ( b ) Quantification of PLA signals per cell as in ( a ). n = 3 independent experiments with n = 10 images per condition and experiment. ( c ) HEK293T cells expressing HA-AVPR1A and FLAG-ACKR3 were lysed (input), the lysate was immunoprecipitated (IP) with anti-HA, followed by immunoblotting (IB) to detect HA-AVPR1A (left) and FLAG-ACKR3 (right) in the IP samples. IP control: precipitate after incubation of cell lysates with IgG-coupled resin. ( d , e ) Intermolecular BRET assays. Cells were co-transfected with AVPR1A-hRluc plus EYFP (open circles) or ACKR3-EYFP (grey squares) at various acceptor : donor ratios ( d ) and with increasing amounts of AVPR1A-hRluc and ACKR3-EYFP at a constant ratio of 1 : 10 ( e ).

    Journal: Open Biology

    Article Title: Identification and functional characterization of arginine vasopressin receptor 1A : atypical chemokine receptor 3 heteromers in vascular smooth muscle

    doi: 10.1098/rsob.170207

    Figure Lengend Snippet: Recombinant ACKR3 and AVPR1A form heteromeric complexes. ( a ) Typical PLA images for the detection of individual receptors and receptor–receptor interactions in HEK293T cells transfected with DNA encoding HA- or FLAG-tagged receptors. Ctrl: cells transfected with pcDNA. Images show merged PLA/DAPI signals acquired from z -stack images ( n = 10; thickness 1 µm, bottom to top). Scale bars, 10 µm. ( b ) Quantification of PLA signals per cell as in ( a ). n = 3 independent experiments with n = 10 images per condition and experiment. ( c ) HEK293T cells expressing HA-AVPR1A and FLAG-ACKR3 were lysed (input), the lysate was immunoprecipitated (IP) with anti-HA, followed by immunoblotting (IB) to detect HA-AVPR1A (left) and FLAG-ACKR3 (right) in the IP samples. IP control: precipitate after incubation of cell lysates with IgG-coupled resin. ( d , e ) Intermolecular BRET assays. Cells were co-transfected with AVPR1A-hRluc plus EYFP (open circles) or ACKR3-EYFP (grey squares) at various acceptor : donor ratios ( d ) and with increasing amounts of AVPR1A-hRluc and ACKR3-EYFP at a constant ratio of 1 : 10 ( e ).

    Article Snippet: A total of 50 µg of rabbit anti-AVPR1A (Bioss BS-11598R), mouse anti-HA (Bioss bsm-50131M) or anti- rabbit IgG (AbCam Ab27478) were incubated with 50 µl of Amino Link Plus coupling resin for 180 min at room temperature.

    Techniques: Recombinant, Proximity Ligation Assay, Transfection, Expressing, Immunoprecipitation, Incubation, Bioluminescence Resonance Energy Transfer

    ACKR3 and AVPR1A form heteromeric complexes in hVSMCs. ( a ) Representative PLA images for the detection of individual receptors, receptor–receptor complexes and pMLC2 in hVSMC. PLA for pMLC2 was performed in non-permeabilized cells (pMLC2, bottom left) and permeabilized cells (pMLC2, permeabilized cells, bottom right). All other PLAs were performed in non-permeabilized cells. Ctrl: omission of one primary antibody. Images show merged PLA/DAPI signals acquired from z -stack images ( n = 10; thickness 1 µm, bottom to top). Scale bars, 10 µm. ( b ) Quantification of PLA signals per cell as in ( a ). n = 4 independent experiments with n = 10 images per condition and experiment. ( c ) Three-dimensional representations of ACKR3 : AVPR1A interactions in hVSMC. Deconvolved images were generated from z -stack images ( n = 20; thickness: 0.5 µm, bottom to top). Images show merged PLA/DAPI signals. ( d–g ) hVSMCs were lysed (=input) and AVPR1A was immunoprecipitated (IP) followed by immunoblotting (IB) to detect AVPR1A ( d ), CXCR4 ( e ), ACKR3 ( f ) and β 2 -AR ( g ) in the IP samples. IP control: precipitate after incubation of cell lysates with IgG-coupled resin. Images are representative of n = 4 independent experiments.

    Journal: Open Biology

    Article Title: Identification and functional characterization of arginine vasopressin receptor 1A : atypical chemokine receptor 3 heteromers in vascular smooth muscle

    doi: 10.1098/rsob.170207

    Figure Lengend Snippet: ACKR3 and AVPR1A form heteromeric complexes in hVSMCs. ( a ) Representative PLA images for the detection of individual receptors, receptor–receptor complexes and pMLC2 in hVSMC. PLA for pMLC2 was performed in non-permeabilized cells (pMLC2, bottom left) and permeabilized cells (pMLC2, permeabilized cells, bottom right). All other PLAs were performed in non-permeabilized cells. Ctrl: omission of one primary antibody. Images show merged PLA/DAPI signals acquired from z -stack images ( n = 10; thickness 1 µm, bottom to top). Scale bars, 10 µm. ( b ) Quantification of PLA signals per cell as in ( a ). n = 4 independent experiments with n = 10 images per condition and experiment. ( c ) Three-dimensional representations of ACKR3 : AVPR1A interactions in hVSMC. Deconvolved images were generated from z -stack images ( n = 20; thickness: 0.5 µm, bottom to top). Images show merged PLA/DAPI signals. ( d–g ) hVSMCs were lysed (=input) and AVPR1A was immunoprecipitated (IP) followed by immunoblotting (IB) to detect AVPR1A ( d ), CXCR4 ( e ), ACKR3 ( f ) and β 2 -AR ( g ) in the IP samples. IP control: precipitate after incubation of cell lysates with IgG-coupled resin. Images are representative of n = 4 independent experiments.

    Article Snippet: A total of 50 µg of rabbit anti-AVPR1A (Bioss BS-11598R), mouse anti-HA (Bioss bsm-50131M) or anti- rabbit IgG (AbCam Ab27478) were incubated with 50 µl of Amino Link Plus coupling resin for 180 min at room temperature.

    Techniques: Proximity Ligation Assay, Generated, Immunoprecipitation, Incubation

    FKN and CX3CR1 mediate WEHI 274.1 recruitment to injured atherosclerotic carotid arteries. ( A ) WEHI 274.1 adhesion to mechanically injured atherosclerotic carotids was studied over 30 minutes in the presence of an IgG control antibody (black bars) or a function blocking anti-FKN antibody (white bars). ( B ) Representative intravital microscopic images from mouse carotid arteries at baseline (left) and 15 minutes (right) after injury of the atherosclerotic vascular wall, pretreated with either an anti-FKN antibody (upper row) or an isotype IgG control antibody (lower row). WEHI 274.1 cells were stained with DCF (green). Bars, 50 µm. n = 4–8, *p

    Journal: PLoS ONE

    Article Title: Fractalkine Is Expressed in Early and Advanced Atherosclerotic Lesions and Supports Monocyte Recruitment via CX3CR1

    doi: 10.1371/journal.pone.0043572

    Figure Lengend Snippet: FKN and CX3CR1 mediate WEHI 274.1 recruitment to injured atherosclerotic carotid arteries. ( A ) WEHI 274.1 adhesion to mechanically injured atherosclerotic carotids was studied over 30 minutes in the presence of an IgG control antibody (black bars) or a function blocking anti-FKN antibody (white bars). ( B ) Representative intravital microscopic images from mouse carotid arteries at baseline (left) and 15 minutes (right) after injury of the atherosclerotic vascular wall, pretreated with either an anti-FKN antibody (upper row) or an isotype IgG control antibody (lower row). WEHI 274.1 cells were stained with DCF (green). Bars, 50 µm. n = 4–8, *p

    Article Snippet: Rat IgG2b and rabbit IgG, used as negative controls, were from Abcam and Dako, respectively ( ).

    Techniques: Blocking Assay, Staining