anti cd11b antibody  (Abcam)

 
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    Name:
    Anti CD11b antibody CD11b
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    Catalog Number:
    AB8879
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    Structured Review

    Abcam anti cd11b antibody
    Masson’s trichrome stain (A, B) and immunofluorescent stain (C–H) of 0 and 7.5 wt % adhesive and surrounding tissue after 2 weeks subcutaneous implantation. a: adhesive. Orange box: cell distribution area. Single headed arrow: cell infiltrating into the pocket formed via gelatin microgel degradation. Blue (DAPI): cell nuclei. Green <t>(CD11b):</t> macrophage. Red (CD163 and S100A4): M2 macrophage and fibroblast, respectively.

    https://www.bioz.com/result/anti cd11b antibody/product/Abcam
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    anti cd11b antibody - by Bioz Stars, 2021-06
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    Images

    1) Product Images from "Gelatin Microgel Incorporated Poly(ethylene glycol)-Based Bioadhesive with Enhanced Adhesive Property and Bioactivity"

    Article Title: Gelatin Microgel Incorporated Poly(ethylene glycol)-Based Bioadhesive with Enhanced Adhesive Property and Bioactivity

    Journal: ACS Applied Materials & Interfaces

    doi: 10.1021/acsami.6b01364

    Masson’s trichrome stain (A, B) and immunofluorescent stain (C–H) of 0 and 7.5 wt % adhesive and surrounding tissue after 2 weeks subcutaneous implantation. a: adhesive. Orange box: cell distribution area. Single headed arrow: cell infiltrating into the pocket formed via gelatin microgel degradation. Blue (DAPI): cell nuclei. Green (CD11b): macrophage. Red (CD163 and S100A4): M2 macrophage and fibroblast, respectively.
    Figure Legend Snippet: Masson’s trichrome stain (A, B) and immunofluorescent stain (C–H) of 0 and 7.5 wt % adhesive and surrounding tissue after 2 weeks subcutaneous implantation. a: adhesive. Orange box: cell distribution area. Single headed arrow: cell infiltrating into the pocket formed via gelatin microgel degradation. Blue (DAPI): cell nuclei. Green (CD11b): macrophage. Red (CD163 and S100A4): M2 macrophage and fibroblast, respectively.

    Techniques Used: Staining

    Masson’s trichrome stain (A, B) and immunofluorescent stain (C–H) of 0 and 7.5 wt % adhesive and surrounding tissue after 6 weeks subcutaneous implantation. a: adhesive. Orange box: cell distribution area. Double headed arrow: collagen layer (CL in A and B). Dashed arrow: cell infiltration layer (IL in A and B). Blue (DAPI): cell nuclei. Green (CD11b): macrophage. Red (CD163 and S100A4): M2 macrophage and fibroblast, respectively.
    Figure Legend Snippet: Masson’s trichrome stain (A, B) and immunofluorescent stain (C–H) of 0 and 7.5 wt % adhesive and surrounding tissue after 6 weeks subcutaneous implantation. a: adhesive. Orange box: cell distribution area. Double headed arrow: collagen layer (CL in A and B). Dashed arrow: cell infiltration layer (IL in A and B). Blue (DAPI): cell nuclei. Green (CD11b): macrophage. Red (CD163 and S100A4): M2 macrophage and fibroblast, respectively.

    Techniques Used: Staining

    2) Product Images from "IL-17 induces macrophages to M2-like phenotype via NF-κB"

    Article Title: IL-17 induces macrophages to M2-like phenotype via NF-κB

    Journal: Cancer Management and Research

    doi: 10.2147/CMAR.S174899

    IL-17 induces CD203 and CD163 expression in mouse peritoneal macrophages via p65. Notes: ( A ) F4/80 and CD11b antibodies were used to authenticate by flow cytometry. ( B and C ) qRT-PCR and immunoreactive bands for p65 in mouse peritoneal macrophages treated with siRNA-p65. ( D ) Flow cytometry analysis of CD206 expression in IL-17-treated macrophages transfected with siRNA-p65. Numerical values denote the mean fluorescence intensity. The red line is the fluorescence intensity of isotype control. The blue line is the fluorescence intensity of CD206. ( E ) Immunofluorescence assay of CD163. Scale bar represents 0.1 mm. ( F ) Bar graph depicts the relative fluorescence intensity of CD163. The data shown represent mean±SD (n=3 per group, *** P
    Figure Legend Snippet: IL-17 induces CD203 and CD163 expression in mouse peritoneal macrophages via p65. Notes: ( A ) F4/80 and CD11b antibodies were used to authenticate by flow cytometry. ( B and C ) qRT-PCR and immunoreactive bands for p65 in mouse peritoneal macrophages treated with siRNA-p65. ( D ) Flow cytometry analysis of CD206 expression in IL-17-treated macrophages transfected with siRNA-p65. Numerical values denote the mean fluorescence intensity. The red line is the fluorescence intensity of isotype control. The blue line is the fluorescence intensity of CD206. ( E ) Immunofluorescence assay of CD163. Scale bar represents 0.1 mm. ( F ) Bar graph depicts the relative fluorescence intensity of CD163. The data shown represent mean±SD (n=3 per group, *** P

    Techniques Used: Expressing, Flow Cytometry, Cytometry, Quantitative RT-PCR, Transfection, Fluorescence, Immunofluorescence

    3) Product Images from "Mitochondrial-Directed Antioxidant Reduces Microglial-Induced Inflammation in Murine In Vitro Model of TC-83 Infection"

    Article Title: Mitochondrial-Directed Antioxidant Reduces Microglial-Induced Inflammation in Murine In Vitro Model of TC-83 Infection

    Journal: Viruses

    doi: 10.3390/v10110606

    Gating strategy for d. ( a ) Uninfected and ( b ) TC-83 infected (MOI:2) BV2 microglia and were collected at 2 hpi and fixed using paraformaldehyde. Cells were stained with phycoerythrin-conjugated Iba1 (PE-A) as a positive control for selection of microglial cells. A phycoerythrin-cyanin5-conjugated CD11b (PE-Cy5-A) antibody was used to determine microglial activation in the context of TC-83 infection. Unstained cells were maintained as a negative control for fluorescence. Figure 3
    Figure Legend Snippet: Gating strategy for d. ( a ) Uninfected and ( b ) TC-83 infected (MOI:2) BV2 microglia and were collected at 2 hpi and fixed using paraformaldehyde. Cells were stained with phycoerythrin-conjugated Iba1 (PE-A) as a positive control for selection of microglial cells. A phycoerythrin-cyanin5-conjugated CD11b (PE-Cy5-A) antibody was used to determine microglial activation in the context of TC-83 infection. Unstained cells were maintained as a negative control for fluorescence. Figure 3

    Techniques Used: Infection, Staining, Positive Control, Selection, Activation Assay, Negative Control, Fluorescence

    4) Product Images from "Deficiency of ITGAM Attenuates Experimental Abdominal Aortic Aneurysm in Mice"

    Article Title: Deficiency of ITGAM Attenuates Experimental Abdominal Aortic Aneurysm in Mice

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    doi: 10.1161/JAHA.120.019900

    CD11b regulates macrophage transendothelial migration. A , Volcano plots show the ‐lg (adjusted P value) against the log 2 (fold change) for each gene. The black horizontal and vertical dashed lines indicate the filtering criteria (|log 2 (fold change)| > 1.5 and P
    Figure Legend Snippet: CD11b regulates macrophage transendothelial migration. A , Volcano plots show the ‐lg (adjusted P value) against the log 2 (fold change) for each gene. The black horizontal and vertical dashed lines indicate the filtering criteria (|log 2 (fold change)| > 1.5 and P

    Techniques Used: Migration

    CD11b does not affect macrophage survival. A , Primary BMDMs from WT and ITGAM(‐/‐) mice were collected and stained with FITC‐Annexin V and PI. The early and late cell apoptosis rates were measured by flow cytometry. No significant difference in the apoptosis rate was observed between the 2 groups (n=3). B , The expression of apoptosis‐related proteins, including p53, caspase3, and Bax, in WT and ITGAM(‐/‐) BMDMs was measured by Western blot. C , A cell viability assay was performed in WT and ITGAM(‐/‐) BMDMs using the Cell Counting Kit‐8. No significant difference in the absorbance (OD 450 nm) was found between the 2 groups (n=3). BMDMs indicates bone marrow–derived macrophages; ITGAM, integrin subunit alpha M; FITC, fluorescein isothiocyanate conjugated; PI, propidium iodide and WT, wild‐type.
    Figure Legend Snippet: CD11b does not affect macrophage survival. A , Primary BMDMs from WT and ITGAM(‐/‐) mice were collected and stained with FITC‐Annexin V and PI. The early and late cell apoptosis rates were measured by flow cytometry. No significant difference in the apoptosis rate was observed between the 2 groups (n=3). B , The expression of apoptosis‐related proteins, including p53, caspase3, and Bax, in WT and ITGAM(‐/‐) BMDMs was measured by Western blot. C , A cell viability assay was performed in WT and ITGAM(‐/‐) BMDMs using the Cell Counting Kit‐8. No significant difference in the absorbance (OD 450 nm) was found between the 2 groups (n=3). BMDMs indicates bone marrow–derived macrophages; ITGAM, integrin subunit alpha M; FITC, fluorescein isothiocyanate conjugated; PI, propidium iodide and WT, wild‐type.

    Techniques Used: Mouse Assay, Staining, Flow Cytometry, Expressing, Western Blot, Viability Assay, Cell Counting, Derivative Assay

    Proposed mechanism of CD11b‐mediated macrophage adhesion and transendothelial migration. (1) Monocyte‐derived CD11b specifically recognizes ICAM on endothelial cells, helping monocytes adhere to endothelial cells and further transmigrate into the subendothelial space. (2) ITGAM deficiency impairs the macrophage transendothelial migratory function by limiting the expression of CCR5, inhibiting the phosphorylation of Akt via RACK1, and reducing the interaction with JAM3. ICAM indicates intercellular adhesion molecule; ITGAM, integrin subunit alpha M; and RACK1, receptor for activated C kinase 1.
    Figure Legend Snippet: Proposed mechanism of CD11b‐mediated macrophage adhesion and transendothelial migration. (1) Monocyte‐derived CD11b specifically recognizes ICAM on endothelial cells, helping monocytes adhere to endothelial cells and further transmigrate into the subendothelial space. (2) ITGAM deficiency impairs the macrophage transendothelial migratory function by limiting the expression of CCR5, inhibiting the phosphorylation of Akt via RACK1, and reducing the interaction with JAM3. ICAM indicates intercellular adhesion molecule; ITGAM, integrin subunit alpha M; and RACK1, receptor for activated C kinase 1.

    Techniques Used: Migration, Derivative Assay, Expressing

    CD11b induces macrophage adhesion. A , Representative images and quantification of WT BMDM adhesion to primary mouse aortic ECs in the absence or presence of CD11b monoclonal antibody or the CD11b agonist LA‐1. The ECs were stimulated with 1 μmol/L angiotensin II overnight before coculture. n=6, * P
    Figure Legend Snippet: CD11b induces macrophage adhesion. A , Representative images and quantification of WT BMDM adhesion to primary mouse aortic ECs in the absence or presence of CD11b monoclonal antibody or the CD11b agonist LA‐1. The ECs were stimulated with 1 μmol/L angiotensin II overnight before coculture. n=6, * P

    Techniques Used:

    CD11b is upregulated in human AAA tissues and a CaCl 2 ‐induced AAA model. A , Representative confocal immunofluorescence staining of CD11b and DAPI in human AAA tissues and control tissues. Scale bars=100 μm. Quantification of CD11b based on the integral optical density in AAA (n=4) and control (n=4) tissues, ** P
    Figure Legend Snippet: CD11b is upregulated in human AAA tissues and a CaCl 2 ‐induced AAA model. A , Representative confocal immunofluorescence staining of CD11b and DAPI in human AAA tissues and control tissues. Scale bars=100 μm. Quantification of CD11b based on the integral optical density in AAA (n=4) and control (n=4) tissues, ** P

    Techniques Used: Immunofluorescence, Staining

    5) Product Images from "The Crotoxin:SBA-15 Complex Down-Regulates the Incidence and Intensity of Experimental Autoimmune Encephalomyelitis Through Peripheral and Central Actions"

    Article Title: The Crotoxin:SBA-15 Complex Down-Regulates the Incidence and Intensity of Experimental Autoimmune Encephalomyelitis Through Peripheral and Central Actions

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2020.591563

    Effect of the treatment with CTX:SBA-15 on neuroinflammation. Animals were treated with CTX (40 µg/kg, s.c.) or CTX:SBA-15 (54 µg/kg, s.c.) in a single administration on the 5 th day after immunization with MOG 35-55 in CFA. The spinal cord of the animals was collected at the peak (A–C) or at 26° after immunization (D) . The cell infiltration analysis was performed by hematoxylin and eosin (HE) staining. Representative sections of cell infiltration are shown (A) as well histological scores (B) of whole spinal cord. Upper (U) and down (D) indicate the magnified corresponding figures. Scale bar 100 μm (n = 4-5) animals per group). The expression of microglia, CD11b (C) , or astrocytes, GFAP (D), markers, were analyzed by Western Blotting assay (n = 6 animals per group). All data are expressed as the mean (± SEM). ***p
    Figure Legend Snippet: Effect of the treatment with CTX:SBA-15 on neuroinflammation. Animals were treated with CTX (40 µg/kg, s.c.) or CTX:SBA-15 (54 µg/kg, s.c.) in a single administration on the 5 th day after immunization with MOG 35-55 in CFA. The spinal cord of the animals was collected at the peak (A–C) or at 26° after immunization (D) . The cell infiltration analysis was performed by hematoxylin and eosin (HE) staining. Representative sections of cell infiltration are shown (A) as well histological scores (B) of whole spinal cord. Upper (U) and down (D) indicate the magnified corresponding figures. Scale bar 100 μm (n = 4-5) animals per group). The expression of microglia, CD11b (C) , or astrocytes, GFAP (D), markers, were analyzed by Western Blotting assay (n = 6 animals per group). All data are expressed as the mean (± SEM). ***p

    Techniques Used: Staining, Expressing, Western Blot

    6) Product Images from "Leukadherin-1-Mediated Activation of CD11b Inhibits LPS-Induced Pro-inflammatory Response in Macrophages and Protects Mice Against Endotoxic Shock by Blocking LPS-TLR4 Interaction"

    Article Title: Leukadherin-1-Mediated Activation of CD11b Inhibits LPS-Induced Pro-inflammatory Response in Macrophages and Protects Mice Against Endotoxic Shock by Blocking LPS-TLR4 Interaction

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2019.00215

    The regulatory mechanism of LA1 on the activation of TLR4 pathway. This schematic shows that activation of CD11b by LA1 induces endocytosis of TLR4 and subsequently, to a certain degree, blocks the interaction of LPS with TLR4 which can decrease activation of TLR4 pathway and inflammation.
    Figure Legend Snippet: The regulatory mechanism of LA1 on the activation of TLR4 pathway. This schematic shows that activation of CD11b by LA1 induces endocytosis of TLR4 and subsequently, to a certain degree, blocks the interaction of LPS with TLR4 which can decrease activation of TLR4 pathway and inflammation.

    Techniques Used: Activation Assay

    Activation of CD11b by LA1 could endocytose TLR4 in macrophages. (A, B) BMDMs were treated with LA1 (20 μM) for 1 and 2 h. FACS analysis was performed to assess the expressions of TLR4 (A) and CD11b (B) in BMDMs. (C, D) C57BL/6 mice were stimulated with LA1 (40 μg/g of body weight) for 1 and 2 h. FACS analysis was performed to assess the expressions of TLR4 (C) and CD11b (D) in PBMCs. (E) Slides containing BMDMs were treated with LA1 (20 μM) for 1 and 2 h, and then, the cells were fixed and permeabilized. The cells were initially stained with anti-TLR4 mAb and anti-CD11b mAb, followed by incubation with an Alexa Fluor 488- and 647-conjugated secondary antibodies. Finally, they were analyzed using a confocal microscope. Red represents CD11b; green represents TLR4; light blue represents nuclei with DAPI; and yellow represents merged TLR4 and CD11b. (F) After treated with LA1 (20 μM) for 1 or 2 h, BMDMs were lysed in a modified RIPA buffer. After entrifuged, one aliquot of the supernatant was saved as the input control, the remainder was separately incubated with anti-CD11b antibody, anti-TLR4 antibody and negative control IgG antibody, followed by pull-down with 30 μl Protein A Agarose beads. The beads were then collected by centrifugation and washed three times with cold PBS. The immunoprecipitates were eluted by boiling in loading buffer and subjected to western blot analysis along with input sample as described above. Data shown are representative of three independent experiments. Error bars represent S.E.M. * p
    Figure Legend Snippet: Activation of CD11b by LA1 could endocytose TLR4 in macrophages. (A, B) BMDMs were treated with LA1 (20 μM) for 1 and 2 h. FACS analysis was performed to assess the expressions of TLR4 (A) and CD11b (B) in BMDMs. (C, D) C57BL/6 mice were stimulated with LA1 (40 μg/g of body weight) for 1 and 2 h. FACS analysis was performed to assess the expressions of TLR4 (C) and CD11b (D) in PBMCs. (E) Slides containing BMDMs were treated with LA1 (20 μM) for 1 and 2 h, and then, the cells were fixed and permeabilized. The cells were initially stained with anti-TLR4 mAb and anti-CD11b mAb, followed by incubation with an Alexa Fluor 488- and 647-conjugated secondary antibodies. Finally, they were analyzed using a confocal microscope. Red represents CD11b; green represents TLR4; light blue represents nuclei with DAPI; and yellow represents merged TLR4 and CD11b. (F) After treated with LA1 (20 μM) for 1 or 2 h, BMDMs were lysed in a modified RIPA buffer. After entrifuged, one aliquot of the supernatant was saved as the input control, the remainder was separately incubated with anti-CD11b antibody, anti-TLR4 antibody and negative control IgG antibody, followed by pull-down with 30 μl Protein A Agarose beads. The beads were then collected by centrifugation and washed three times with cold PBS. The immunoprecipitates were eluted by boiling in loading buffer and subjected to western blot analysis along with input sample as described above. Data shown are representative of three independent experiments. Error bars represent S.E.M. * p

    Techniques Used: Activation Assay, FACS, Mouse Assay, Staining, Incubation, Microscopy, Modification, Negative Control, Centrifugation, Western Blot

    Activation of CD11b by LA1 alleviated LPS-induced lung and liver injury. (A, B) C57BL/6 mice were treated with either LA1 (10, 20, and 40 μg/g of body weight) or vehicle for 2 h, and then, stimulated with LPS (10 μg/g of body weight). After 12 h, tissues of lung and liver were fixed with 4% paraformaldehyde and paraffin-embedded lung (A) and liver (B) sections were stained with H E. (C, D) C57BL/6 mice were treated with either LA1 (40 μg/g of body weight) or vehicle for 2 h, and then, stimulated with LPS (10 μg/g of body weight). After 12 h, apoptosis in the cells of lung (C) and liver (D) was analyzed by TUNEL. Data shown are representative of three independent experiments. Error bars represent S.E.M. * p
    Figure Legend Snippet: Activation of CD11b by LA1 alleviated LPS-induced lung and liver injury. (A, B) C57BL/6 mice were treated with either LA1 (10, 20, and 40 μg/g of body weight) or vehicle for 2 h, and then, stimulated with LPS (10 μg/g of body weight). After 12 h, tissues of lung and liver were fixed with 4% paraformaldehyde and paraffin-embedded lung (A) and liver (B) sections were stained with H E. (C, D) C57BL/6 mice were treated with either LA1 (40 μg/g of body weight) or vehicle for 2 h, and then, stimulated with LPS (10 μg/g of body weight). After 12 h, apoptosis in the cells of lung (C) and liver (D) was analyzed by TUNEL. Data shown are representative of three independent experiments. Error bars represent S.E.M. * p

    Techniques Used: Activation Assay, Mouse Assay, Staining, TUNEL Assay

    Activation of CD11b by LA1 inhibited LPS-induced pro-inflammatory response in macrophages in vivo . C57BL/6 mice were treated with either LA1 (10, 20, and 40 μg/g of body weight) or vehicle for 2 h, and then, stimulated with LPS (10 μg/g of body weight). FACS analysis was performed to assess the expressions of CD86 (A) and CD40 (B) in splenic macrophages after 12 h. Levels of IL-6, TNF-α, IL-12, and IL-1β in serum were determined by ELISA after 6 h (C) . The expressions of IL-6, TNF-α, IL-12, and IL-1β (D) in peritoneal cells were examined by qRT-PCR after 6 h. Data are shown as the means ± SEM ( n = 6 replicates) and are representative of three independent experiments. Error bars represent S.E.M. * p
    Figure Legend Snippet: Activation of CD11b by LA1 inhibited LPS-induced pro-inflammatory response in macrophages in vivo . C57BL/6 mice were treated with either LA1 (10, 20, and 40 μg/g of body weight) or vehicle for 2 h, and then, stimulated with LPS (10 μg/g of body weight). FACS analysis was performed to assess the expressions of CD86 (A) and CD40 (B) in splenic macrophages after 12 h. Levels of IL-6, TNF-α, IL-12, and IL-1β in serum were determined by ELISA after 6 h (C) . The expressions of IL-6, TNF-α, IL-12, and IL-1β (D) in peritoneal cells were examined by qRT-PCR after 6 h. Data are shown as the means ± SEM ( n = 6 replicates) and are representative of three independent experiments. Error bars represent S.E.M. * p

    Techniques Used: Activation Assay, In Vivo, Mouse Assay, FACS, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

    Activation of CD11b by LA1 inhibited LPS/IFN-γ-induced pro-inflammatory response in macrophages in vitro . (A–D) BMDMs were pretreated with LA1 (0, 5, 10, and 20 μM) for 2 h, and then, the cells were treated with LPS (200 ng/ml) and IFN-γ (10 ng/ml). FACS analysis was performed to assess the expressions of CD86 (A) and CD40 (B) after 24 h. Levels of IL-6, TNF-α, IL-12, and IL-1β in supernatant were determined by ELISA after 24 h (C) . Expressions of IL-6, TNF-α, IL-12, and IL-1β were measured using qRT-PCR after 6 h (D) . (E) BMDMs were pretreated with either 20 μM LA1 or vehicle for 2 h, and then, the cells were stimulated with LPS (200 ng/ml) and IFN-γ (10 ng/ml) for 30 and 60 min. Cell lysates were prepared and subjected to immunoblotting with the indicated antibodies. β-actin was used as loading control. Data are representative from one out of three biological replicates, each with three technical replicates. Error bars represent S.E.M. * p
    Figure Legend Snippet: Activation of CD11b by LA1 inhibited LPS/IFN-γ-induced pro-inflammatory response in macrophages in vitro . (A–D) BMDMs were pretreated with LA1 (0, 5, 10, and 20 μM) for 2 h, and then, the cells were treated with LPS (200 ng/ml) and IFN-γ (10 ng/ml). FACS analysis was performed to assess the expressions of CD86 (A) and CD40 (B) after 24 h. Levels of IL-6, TNF-α, IL-12, and IL-1β in supernatant were determined by ELISA after 24 h (C) . Expressions of IL-6, TNF-α, IL-12, and IL-1β were measured using qRT-PCR after 6 h (D) . (E) BMDMs were pretreated with either 20 μM LA1 or vehicle for 2 h, and then, the cells were stimulated with LPS (200 ng/ml) and IFN-γ (10 ng/ml) for 30 and 60 min. Cell lysates were prepared and subjected to immunoblotting with the indicated antibodies. β-actin was used as loading control. Data are representative from one out of three biological replicates, each with three technical replicates. Error bars represent S.E.M. * p

    Techniques Used: Activation Assay, In Vitro, FACS, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

    Activation of TLR4 by LPS could endocytose CD11b in macrophages. (A, B) BMDMs were treated with LPS (200 ng/ml) and IFN-γ (10 ng/ml) for 1, 2, and 4 h. FACS analysis was performed to assess the expressions of TLR4 (A) and CD11b (B) in BMDMs. (C–F) C57BL/6 mice were stimulated by LPS (10 μg/g of body weight) for 1, 2, and 4 h. FACS analysis was performed to assess the expressions of TLR4 (C) and CD11b (D) in peritoneal cavity, as well as to assess the expressions of TLR4 (E) and CD11b (F) in PBMCs. (G) BMDMs containing slides were treated with LPS (200 ng/ml) and IFN-γ (10 ng/ml) for 1, 2, and 4 h, and then, the cells were fixed and permeabilized. After staining with anti-TLR4 mAb and anti-CD11b mAb, cells were incubated with an Alexa Fluor 488- and 647-conjugated secondary antibody and analyzed by fluorescent staining. Red represents CD11b; green represents TLR4; light blue represents nuclei with DAPI; and yellow represents merged TLR4 and CD11b. For in vitro studies, the data represent one out of the three biological replicates, each with three technical replicates. The data are shown as the means ± SEM ( n = 6 replicates) and are representative of three independent experiments. Error bars represent S.E.M. * p
    Figure Legend Snippet: Activation of TLR4 by LPS could endocytose CD11b in macrophages. (A, B) BMDMs were treated with LPS (200 ng/ml) and IFN-γ (10 ng/ml) for 1, 2, and 4 h. FACS analysis was performed to assess the expressions of TLR4 (A) and CD11b (B) in BMDMs. (C–F) C57BL/6 mice were stimulated by LPS (10 μg/g of body weight) for 1, 2, and 4 h. FACS analysis was performed to assess the expressions of TLR4 (C) and CD11b (D) in peritoneal cavity, as well as to assess the expressions of TLR4 (E) and CD11b (F) in PBMCs. (G) BMDMs containing slides were treated with LPS (200 ng/ml) and IFN-γ (10 ng/ml) for 1, 2, and 4 h, and then, the cells were fixed and permeabilized. After staining with anti-TLR4 mAb and anti-CD11b mAb, cells were incubated with an Alexa Fluor 488- and 647-conjugated secondary antibody and analyzed by fluorescent staining. Red represents CD11b; green represents TLR4; light blue represents nuclei with DAPI; and yellow represents merged TLR4 and CD11b. For in vitro studies, the data represent one out of the three biological replicates, each with three technical replicates. The data are shown as the means ± SEM ( n = 6 replicates) and are representative of three independent experiments. Error bars represent S.E.M. * p

    Techniques Used: Activation Assay, FACS, Mouse Assay, Staining, Incubation, In Vitro

    Activation of CD11b by LA1 reduced LPS-induced mortality. (A) C57BL/6 mice were treated with either LA1 (40 μg/g of body weight) or vehicle followed by LPS stimulation (25, 37.5, and 50 μg/g of body weight). The survivals of mice were observed ( n = 10 mice/group) ** P
    Figure Legend Snippet: Activation of CD11b by LA1 reduced LPS-induced mortality. (A) C57BL/6 mice were treated with either LA1 (40 μg/g of body weight) or vehicle followed by LPS stimulation (25, 37.5, and 50 μg/g of body weight). The survivals of mice were observed ( n = 10 mice/group) ** P

    Techniques Used: Activation Assay, Mouse Assay

    7) Product Images from "The Potential of the FSP1cre-Pparb/d−/− Mouse Model for Studying Juvenile NAFLD"

    Article Title: The Potential of the FSP1cre-Pparb/d−/− Mouse Model for Studying Juvenile NAFLD

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20205115

    Distribution of FSP1 + CD11b + and FSP1 + CD11b − cells in non-parenchymal liver cells and genotyping of these isolated cells. Majority of the FSP1 + cells express CD11b. ( a ) Pparb/d fl/fl and FSP1cre- Pparb/d −/− liver FACS plots showing the CD45 + cell population, which is stained for FSP1 and CD11b, and graphs showing the distribution of cells from each quadrant in percentage (data presented as mean ± s.e.m; n = 6 biological replicates per group). Two-tailed Mann-Whitney test of the data did not find significant differences between the FSP1cre- Pparb/d −/− and Pparb/d fl/fl genotypes. ( b ) Schematic diagram showing the relative position of PCR genotyping primers for the Pparb/d lox/lox (400 bp band) and the deleted Pparb/d exon 4 alleles (490 bp band). FSP1-cre genotyping: 100 bp band indicates FSP1-cre. 300 bp band is from the endogenous Fsp1 gene. In both gels, the rightmost lane depicts the molecular weight 100 bp ladder. ( c ) Fsp1 and Pparb/d gene expression in isolated FSP1 + CD11b + cells from P2 Pparb/d fl/fl and FSP1cre- Pparb/d −/− livers. BD, below detection. Two-tailed Mann-Whitney test of the data shown as mean ± s.e.m, n = 5 biological replicates per group. ** P
    Figure Legend Snippet: Distribution of FSP1 + CD11b + and FSP1 + CD11b − cells in non-parenchymal liver cells and genotyping of these isolated cells. Majority of the FSP1 + cells express CD11b. ( a ) Pparb/d fl/fl and FSP1cre- Pparb/d −/− liver FACS plots showing the CD45 + cell population, which is stained for FSP1 and CD11b, and graphs showing the distribution of cells from each quadrant in percentage (data presented as mean ± s.e.m; n = 6 biological replicates per group). Two-tailed Mann-Whitney test of the data did not find significant differences between the FSP1cre- Pparb/d −/− and Pparb/d fl/fl genotypes. ( b ) Schematic diagram showing the relative position of PCR genotyping primers for the Pparb/d lox/lox (400 bp band) and the deleted Pparb/d exon 4 alleles (490 bp band). FSP1-cre genotyping: 100 bp band indicates FSP1-cre. 300 bp band is from the endogenous Fsp1 gene. In both gels, the rightmost lane depicts the molecular weight 100 bp ladder. ( c ) Fsp1 and Pparb/d gene expression in isolated FSP1 + CD11b + cells from P2 Pparb/d fl/fl and FSP1cre- Pparb/d −/− livers. BD, below detection. Two-tailed Mann-Whitney test of the data shown as mean ± s.e.m, n = 5 biological replicates per group. ** P

    Techniques Used: Isolation, FACS, Staining, Two Tailed Test, MANN-WHITNEY, Polymerase Chain Reaction, Molecular Weight, Expressing

    Double immunofluorescence staining in Fibroblast-specific protein 1 (FSP1)cre- Pparb/d −/− liver for Glial fibrillary acidic protein (GFAP), CD146, CD68 or CD11b with FSP1. ( a ) CD11b-expressing liver resident macrophages express FSP1. ( b ) CD68-expressing Kupffer cells do not express FSP1. ( c ) GFAP-expressing hepatic stellate cells (HSCs) do not express FSP1. ( d ) CD146-expressing LSECs do not express FSP1. Arrows indicate staining; the green arrow indicates positive co-staining of CD11b and FSP1.
    Figure Legend Snippet: Double immunofluorescence staining in Fibroblast-specific protein 1 (FSP1)cre- Pparb/d −/− liver for Glial fibrillary acidic protein (GFAP), CD146, CD68 or CD11b with FSP1. ( a ) CD11b-expressing liver resident macrophages express FSP1. ( b ) CD68-expressing Kupffer cells do not express FSP1. ( c ) GFAP-expressing hepatic stellate cells (HSCs) do not express FSP1. ( d ) CD146-expressing LSECs do not express FSP1. Arrows indicate staining; the green arrow indicates positive co-staining of CD11b and FSP1.

    Techniques Used: Double Immunofluorescence Staining, Expressing, Staining

    8) Product Images from "Exosomes secreted by prostate cancer cells under hypoxia promote matrix metalloproteinases activity at pre-metastatic niches"

    Article Title: Exosomes secreted by prostate cancer cells under hypoxia promote matrix metalloproteinases activity at pre-metastatic niches

    Journal: Molecular carcinogenesis

    doi: 10.1002/mc.23157

    Effect of PCa Exo Hypoxic treatment on CD11b+ cells in male athymic nude mice. Male athymic nude mice were injected with PBS, Exo Normoxic or Exo Hypoxic (10 μg each, IP) on alternate days for 4 weeks. At the end, mice were killed and mentioned organs were collected, processed, and analyzed for CD11b+ cells by IHC and presented as % positive cells. Data shown in the bar diagram represent mean ± SEM of immunoreactivity scores for ten randomly selected fields from 4 to 5 samples for each group. Representative images are shown at ×400. * P ≤ .05. Exo Hypoxic , exosomes secreted under hypoxic condition; Exo Normoxic ]
    Figure Legend Snippet: Effect of PCa Exo Hypoxic treatment on CD11b+ cells in male athymic nude mice. Male athymic nude mice were injected with PBS, Exo Normoxic or Exo Hypoxic (10 μg each, IP) on alternate days for 4 weeks. At the end, mice were killed and mentioned organs were collected, processed, and analyzed for CD11b+ cells by IHC and presented as % positive cells. Data shown in the bar diagram represent mean ± SEM of immunoreactivity scores for ten randomly selected fields from 4 to 5 samples for each group. Representative images are shown at ×400. * P ≤ .05. Exo Hypoxic , exosomes secreted under hypoxic condition; Exo Normoxic ]

    Techniques Used: Mouse Assay, Injection, Immunohistochemistry

    9) Product Images from "Retinal Detachment-Induced Müller Glial Cell Swelling Activates TRPV4 Ion Channels and Triggers Photoreceptor Death at Body Temperature"

    Article Title: Retinal Detachment-Induced Müller Glial Cell Swelling Activates TRPV4 Ion Channels and Triggers Photoreceptor Death at Body Temperature

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0897-18.2018

    RD-induced TRPV4 activation recruits macrophages in subretinal space. Immunostaining of CD11b (a macrophage marker, represented as green) was performed in RD retinae. Nuclei were stained by Hoechst (blue). Shown is quantification of CD11b-positive cell numbers in WT or TRPV4 retinae 24 h after retinal detachments ( n = 8, ** p = 0.00632, Student t test). Scale bar, 100 μm. ONL, Outer nuclear layer; INL, inner nuclear layer.
    Figure Legend Snippet: RD-induced TRPV4 activation recruits macrophages in subretinal space. Immunostaining of CD11b (a macrophage marker, represented as green) was performed in RD retinae. Nuclei were stained by Hoechst (blue). Shown is quantification of CD11b-positive cell numbers in WT or TRPV4 retinae 24 h after retinal detachments ( n = 8, ** p = 0.00632, Student t test). Scale bar, 100 μm. ONL, Outer nuclear layer; INL, inner nuclear layer.

    Techniques Used: Activation Assay, Immunostaining, Marker, Staining

    10) Product Images from "Overexpression of miR-146a Might Regulate Polarization Transitions of BV-2 Cells Induced by High Glucose and Glucose Fluctuations"

    Article Title: Overexpression of miR-146a Might Regulate Polarization Transitions of BV-2 Cells Induced by High Glucose and Glucose Fluctuations

    Journal: Frontiers in Endocrinology

    doi: 10.3389/fendo.2019.00719

    Immunofluorescence for CD11b in BV-2 cells overexpressing miR-146a induced by high glucose and glucose fluctuations for 48 h. (A) Immunofluorescence was used to measure the expression of CD11b protein in BV-2 cells overexpressing miR-146a induced by high glucose and glucose fluctuation. (B) Comparison of CD11b protein immunofluorescence intensity. ### p
    Figure Legend Snippet: Immunofluorescence for CD11b in BV-2 cells overexpressing miR-146a induced by high glucose and glucose fluctuations for 48 h. (A) Immunofluorescence was used to measure the expression of CD11b protein in BV-2 cells overexpressing miR-146a induced by high glucose and glucose fluctuation. (B) Comparison of CD11b protein immunofluorescence intensity. ### p

    Techniques Used: Immunofluorescence, Expressing

    Effects of miR-146a on polarization transitions in BV-2 cells induced by high glucose and glucose fluctuation. (A) CD11b mRNA expression levels. *** p
    Figure Legend Snippet: Effects of miR-146a on polarization transitions in BV-2 cells induced by high glucose and glucose fluctuation. (A) CD11b mRNA expression levels. *** p

    Techniques Used: Expressing

    11) Product Images from "Microglia Polarization with M1/M2 Phenotype Changes in rd1 Mouse Model of Retinal Degeneration"

    Article Title: Microglia Polarization with M1/M2 Phenotype Changes in rd1 Mouse Model of Retinal Degeneration

    Journal: Frontiers in Neuroanatomy

    doi: 10.3389/fnana.2017.00077

    Activated microglia skew to M1 polarization during the retinal degeneration of rd1 mouse. (A) Although the gating of alive cells reduced in rd1 retinae comparing with C57 mice, the percentage of microglia increased significantly in rd1 mice. Microglia were identified as CD45 int CD11b + cells (P14: 11.4767 ± 1.07653, p = 0.00043; P21:7.92667 ± 0.615639, p = 0.0002; P28: 11.7967 ± 0.27302, p
    Figure Legend Snippet: Activated microglia skew to M1 polarization during the retinal degeneration of rd1 mouse. (A) Although the gating of alive cells reduced in rd1 retinae comparing with C57 mice, the percentage of microglia increased significantly in rd1 mice. Microglia were identified as CD45 int CD11b + cells (P14: 11.4767 ± 1.07653, p = 0.00043; P21:7.92667 ± 0.615639, p = 0.0002; P28: 11.7967 ± 0.27302, p

    Techniques Used: Mouse Assay

    Related Articles

    Immunofluorescence:

    Article Title: Glaucomatous Tissue Stress and the Regulation of Immune Response through Glial Toll-like Receptor Signaling
    Article Snippet: All procedures were similar to those published., , , Monoclonal antibodies to TLR2, -3, and -4 (2 μg/mL; Assay Designs, Ann Arbor, MI) served as the primary antibodies. .. In addition, antibodies against CD11b (1:100; Abcam, Cambridge, MA) or glial fibrillary acidic protein (GFAP; 1:200; Santa Cruz Biotechnology, Santa Cruz, CA) were used to identify microglia and astrocytes during double immunofluorescence labeling. .. A mixture of Alexa Fluor 488- or 568-conjugated species-specific IgGs (1:400; Molecular Probes, Eugene, OR) was used for the secondary antibody incubation.

    Article Title: Organoid modeling of the tumor immune microenvironment
    Article Snippet: In order to remove common sequencing artifacts or residual germline variation, each mutation in the combined MAF file was subjected to a “Panel of Normals” filtering using a panel of over 4000 BAM files from normal samples. .. Mouse immunofluorescence analysis utilized anti-Mouse CD8a (4SM16, BD, 1:100), anti-mouse CD3e (145–2C11, BD, 1:100), anti-mouse CD4 (4SM95, BD, 1:100), anti-CD11b (ab1333357, Abcam, 1:2000), anti-Vimentin (Ab5733, EMD, 1:2000), anti-CD20(Abcam, 1:100). .. All images were captured on a Zeiss Axio-Imager Z1 with ApoTome attachment.

    Labeling:

    Article Title: Glaucomatous Tissue Stress and the Regulation of Immune Response through Glial Toll-like Receptor Signaling
    Article Snippet: All procedures were similar to those published., , , Monoclonal antibodies to TLR2, -3, and -4 (2 μg/mL; Assay Designs, Ann Arbor, MI) served as the primary antibodies. .. In addition, antibodies against CD11b (1:100; Abcam, Cambridge, MA) or glial fibrillary acidic protein (GFAP; 1:200; Santa Cruz Biotechnology, Santa Cruz, CA) were used to identify microglia and astrocytes during double immunofluorescence labeling. .. A mixture of Alexa Fluor 488- or 568-conjugated species-specific IgGs (1:400; Molecular Probes, Eugene, OR) was used for the secondary antibody incubation.

    Incubation:

    Article Title: Human-induced pluripotent stem cell-derived macrophages and their immunological function in response to tuberculosis infection
    Article Snippet: .. They were then incubated at 4 °C overnight with the following antibodies: rabbit anti-CD11b (1:250; Abcam, Cambridge, MA USA), rabbit anti-CD14(1:500; PeproTech), rabbit anti-CD68 (1:500; PeproTech), rabbit anti-MHC class II(1;250; Abcam). .. The cells were washed three times with PBS and then incubated with the following secondary antibodies: Alexa Fluor 555-labeled donkey anti-rabbit IgG (H + L) (1:500; Beyotime), nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (1 μg/ml; Beyotime).

    Article Title: The Role of the Intestinal Microbiome in Chronic Psychosocial Stress-Induced Pathologies in Male Mice
    Article Snippet: .. Following visualization and digitalization, membranes were again stripped and this time incubated with CD11b antibody (1:2000; Abcam® , Cambridge, United Kingdom) over night at 4°C. .. Following incubation with HRP-conjugated anti-rabbit antibody (1:2000), visualization and digitalization were again performed as described above.

    Staining:

    Article Title: LDL receptor-related protein-1 regulates NFκB and microRNA-155 in macrophages to control the inflammatory response
    Article Snippet: Four-micrometer-thick tissue sections were cut and mounted on positively charged microscope slides. .. Sections were stained with antibody against CD11b (1:1,200) from Abcam using a Ventana Discovery Ultra (Ventana Medical Systems). ..

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    Abcam monoclonal antibodies targeting cd11b
    Photographs (40 × magnification) illustrating the effect of nefopam on spinal immunoreactivity to cluster of differentiation molecule 11b <t>(CD11b)</t> after left L5 spinal nerve ligation. The immunohistochemical intensity of CD11b in the L5 segment decreased in a dose dependent manner following nefopam administration. ( A ) sham surgery group, ( B ) normal saline group, ( C ) 10 µg/kg of nefopam group, ( D ) 100 µg/kg of nefopam group.
    Monoclonal Antibodies Targeting Cd11b, 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
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    Abcam rabbit anti cd11b antibody
    Attenuated TDC in vessel wall of ASO and diabetic foot patients compared to healthy person (200 ×). A. Representative images of <t>CD11b-expressing</t> TDC (left) and DC-SIGN-expressing TDC (right) in the vessel wall of healthy person. B. Representative
    Rabbit Anti Cd11b Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cd11b  (Abcam)
    96
    Abcam cd11b
    Mass spectrometric and immunohistochemical staining of thioglycolate-elicited peritoneal cells (eMPCs), polarized macrophages, and DCs. Panels A–B: Hierarchical cluster analysis of eMPCs. Cells were harvested from the peritoneal cavity of C57BL/6J- Ldlr tm1Her mice 5 days after intraperitoneal injection with thioglycolate. Isolated plasma membrane proteins detected by LC-MS/MS analysis of eMPCs were subjected to hierarchical cluster analysis, using the 107 proteins identified as differentially expressed by myeloid cells generated in vitro ( Fig. 2 ). Panel B: Relationships among eMPCs, M1 cells, M2 cells, BmMs, and DCs, as determined by cluster analysis. Panel C: Protein expression in eMPCs, M1 cells, M2 cells, BmMs, and BmDCs. Protein levels were quantified by MS/MS and spectral counting. Data are presented as means and SDs. Panels D–E: Flow cytometric analysis of <t>CD11b,</t> CD11c, and F4/80 in eMPCs. Results are presented as contour plots with 10% probability increments. Panel F: qRT-PCR analysis of M1 marker genes ( Nos2 , Il12b , Tnfa ) in M1 macrophages, BmDCs, and eMPCs. Results (means and SEMs; N = 6) were standardized to 18S levels and expressed relative to M1 macrophages. Panel G: Immunostaining of eMPCs. Cells were stained with antibodies (red channel) to plasma membrane proteins differentially expressed by BmDCs (MBC2, FER1L), BmMs (ITGA6, STAB1), M2 cells (TFRC, ITGB5), and M1 cells (CD11a, CD40). Nuclei were visualized by DAPI staining (blue channel). Immunostaining and microscopy were performed on the same day and with identical microscope settings to experiments presented in Fig. 3D and Fig. S3 . Results obtained for flow cytometry, qRT-PCR, and immunocotyochemistry are representative of 3 independent analyses.
    Cd11b, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Photographs (40 × magnification) illustrating the effect of nefopam on spinal immunoreactivity to cluster of differentiation molecule 11b (CD11b) after left L5 spinal nerve ligation. The immunohistochemical intensity of CD11b in the L5 segment decreased in a dose dependent manner following nefopam administration. ( A ) sham surgery group, ( B ) normal saline group, ( C ) 10 µg/kg of nefopam group, ( D ) 100 µg/kg of nefopam group.

    Journal: Journal of Korean Medical Science

    Article Title: Mechanical Antiallodynic Effect of Intrathecal Nefopam in a Rat Neuropathic Pain Model

    doi: 10.3346/jkms.2015.30.8.1189

    Figure Lengend Snippet: Photographs (40 × magnification) illustrating the effect of nefopam on spinal immunoreactivity to cluster of differentiation molecule 11b (CD11b) after left L5 spinal nerve ligation. The immunohistochemical intensity of CD11b in the L5 segment decreased in a dose dependent manner following nefopam administration. ( A ) sham surgery group, ( B ) normal saline group, ( C ) 10 µg/kg of nefopam group, ( D ) 100 µg/kg of nefopam group.

    Article Snippet: In the present study, monoclonal antibodies targeting CD11b (ab52478, Abcam plc., Cambridge, MA, USA) and GFAP (ab7260, Abcam plc., Cambridge, MA, USA) were used as primary antibodies.

    Techniques: Ligation, Immunohistochemistry

    Changes in the relative mean optical density (MOD) ratio for ( A ) CD11b and ( B ) GFAP after left L5 spinal nerve ligation. The relative MOD ratio increased in the spinal cord side ipsilateral to the spinal nerve ligation in rats. The relative MOD ratio decreased in a dose-dependent manner in rats administered intrathecal nefopam (N10 and N100) compared to the ratio in rats administered intrathecal saline (group C). * P

    Journal: Journal of Korean Medical Science

    Article Title: Mechanical Antiallodynic Effect of Intrathecal Nefopam in a Rat Neuropathic Pain Model

    doi: 10.3346/jkms.2015.30.8.1189

    Figure Lengend Snippet: Changes in the relative mean optical density (MOD) ratio for ( A ) CD11b and ( B ) GFAP after left L5 spinal nerve ligation. The relative MOD ratio increased in the spinal cord side ipsilateral to the spinal nerve ligation in rats. The relative MOD ratio decreased in a dose-dependent manner in rats administered intrathecal nefopam (N10 and N100) compared to the ratio in rats administered intrathecal saline (group C). * P

    Article Snippet: In the present study, monoclonal antibodies targeting CD11b (ab52478, Abcam plc., Cambridge, MA, USA) and GFAP (ab7260, Abcam plc., Cambridge, MA, USA) were used as primary antibodies.

    Techniques: Ligation

    Attenuated TDC in vessel wall of ASO and diabetic foot patients compared to healthy person (200 ×). A. Representative images of CD11b-expressing TDC (left) and DC-SIGN-expressing TDC (right) in the vessel wall of healthy person. B. Representative

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Characteristics of immunogenic and tolerogenic dendritic cells within the arterial wall in atherosclerosis and in vitro

    doi:

    Figure Lengend Snippet: Attenuated TDC in vessel wall of ASO and diabetic foot patients compared to healthy person (200 ×). A. Representative images of CD11b-expressing TDC (left) and DC-SIGN-expressing TDC (right) in the vessel wall of healthy person. B. Representative

    Article Snippet: Rabbit anti-CD11b antibody (ab133571) was obtained from Abcom; Rabbit anti-IDO antibody (bs-2379R) and Rabbit anti-DC-SIGN (bs-2557R) were obtained from Bioss (Beijing, China); Rabbit anti-CD83 (LS-C176864) antibody was obtained from LSBio.

    Techniques: Allele-specific Oligonucleotide, Expressing

    CD11c + CD11b + TDC and CD11c + CD83 + mDC were induced successfully and displayed tolerogenic phenotype. A. Left dots plot showed that the ratio of rmGM-CSF and IL-4 induced CD11c + CD11b + TDC was approximately 41.5%, and right dots plot showed that the ratio of

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Characteristics of immunogenic and tolerogenic dendritic cells within the arterial wall in atherosclerosis and in vitro

    doi:

    Figure Lengend Snippet: CD11c + CD11b + TDC and CD11c + CD83 + mDC were induced successfully and displayed tolerogenic phenotype. A. Left dots plot showed that the ratio of rmGM-CSF and IL-4 induced CD11c + CD11b + TDC was approximately 41.5%, and right dots plot showed that the ratio of

    Article Snippet: Rabbit anti-CD11b antibody (ab133571) was obtained from Abcom; Rabbit anti-IDO antibody (bs-2379R) and Rabbit anti-DC-SIGN (bs-2557R) were obtained from Bioss (Beijing, China); Rabbit anti-CD83 (LS-C176864) antibody was obtained from LSBio.

    Techniques:

    Quantification analyzed the expression of TDC and mDC in ASO and diabetic foot patients. A. Western blotting analyzed the expression of CD1a and CD83 in TDC and CD11b and DC-SIGN in mDC of ASO and diabetic foot patients. B. Quantification of the protein

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Characteristics of immunogenic and tolerogenic dendritic cells within the arterial wall in atherosclerosis and in vitro

    doi:

    Figure Lengend Snippet: Quantification analyzed the expression of TDC and mDC in ASO and diabetic foot patients. A. Western blotting analyzed the expression of CD1a and CD83 in TDC and CD11b and DC-SIGN in mDC of ASO and diabetic foot patients. B. Quantification of the protein

    Article Snippet: Rabbit anti-CD11b antibody (ab133571) was obtained from Abcom; Rabbit anti-IDO antibody (bs-2379R) and Rabbit anti-DC-SIGN (bs-2557R) were obtained from Bioss (Beijing, China); Rabbit anti-CD83 (LS-C176864) antibody was obtained from LSBio.

    Techniques: Expressing, Allele-specific Oligonucleotide, Western Blot

    Effects of fecal transplantation (FT) on local colonic inflammatory parameters. The inflammatory status locally in the colon of single-housed control (SHC) and chronic subordinate colony (CSC) housing mice was assessed at the histological, protein and mRNA level. There was no effect of CSC and/or FT on the histological damage score (A) , the colonic protein levels of F4/80 (B) and cluster of differentiation molecule 11b (CD11b; C ) as well as the mRNA levels of tumor necrosis factor-α (TNFα; D ), interferon-γ (IFNγ; E ) and cathelin-related antimicrobial peptide (CRAMP; F ) in colonic tissue. SHC ( n = 7–19); CSC ( n = 4–13). Parametric data are presented as mean + SEM. Non-parametric data are represented as box-plots. Solid line represents the median, dotted line represents the mean for each data set. Lower boxes indicate 25th, upper boxes indicate 75th percentile, 10th (lower error bar), and 90th percentile (upper error bar) as well as possible outliers beyond the percentiles (indicated by closed circles) are also shown.

    Journal: Frontiers in Behavioral Neuroscience

    Article Title: The Role of the Intestinal Microbiome in Chronic Psychosocial Stress-Induced Pathologies in Male Mice

    doi: 10.3389/fnbeh.2018.00252

    Figure Lengend Snippet: Effects of fecal transplantation (FT) on local colonic inflammatory parameters. The inflammatory status locally in the colon of single-housed control (SHC) and chronic subordinate colony (CSC) housing mice was assessed at the histological, protein and mRNA level. There was no effect of CSC and/or FT on the histological damage score (A) , the colonic protein levels of F4/80 (B) and cluster of differentiation molecule 11b (CD11b; C ) as well as the mRNA levels of tumor necrosis factor-α (TNFα; D ), interferon-γ (IFNγ; E ) and cathelin-related antimicrobial peptide (CRAMP; F ) in colonic tissue. SHC ( n = 7–19); CSC ( n = 4–13). Parametric data are presented as mean + SEM. Non-parametric data are represented as box-plots. Solid line represents the median, dotted line represents the mean for each data set. Lower boxes indicate 25th, upper boxes indicate 75th percentile, 10th (lower error bar), and 90th percentile (upper error bar) as well as possible outliers beyond the percentiles (indicated by closed circles) are also shown.

    Article Snippet: Following visualization and digitalization, membranes were again stripped and this time incubated with CD11b antibody (1:2000; Abcam® , Cambridge, United Kingdom) over night at 4°C.

    Techniques: Transplantation Assay, Mouse Assay

    Mass spectrometric and immunohistochemical staining of thioglycolate-elicited peritoneal cells (eMPCs), polarized macrophages, and DCs. Panels A–B: Hierarchical cluster analysis of eMPCs. Cells were harvested from the peritoneal cavity of C57BL/6J- Ldlr tm1Her mice 5 days after intraperitoneal injection with thioglycolate. Isolated plasma membrane proteins detected by LC-MS/MS analysis of eMPCs were subjected to hierarchical cluster analysis, using the 107 proteins identified as differentially expressed by myeloid cells generated in vitro ( Fig. 2 ). Panel B: Relationships among eMPCs, M1 cells, M2 cells, BmMs, and DCs, as determined by cluster analysis. Panel C: Protein expression in eMPCs, M1 cells, M2 cells, BmMs, and BmDCs. Protein levels were quantified by MS/MS and spectral counting. Data are presented as means and SDs. Panels D–E: Flow cytometric analysis of CD11b, CD11c, and F4/80 in eMPCs. Results are presented as contour plots with 10% probability increments. Panel F: qRT-PCR analysis of M1 marker genes ( Nos2 , Il12b , Tnfa ) in M1 macrophages, BmDCs, and eMPCs. Results (means and SEMs; N = 6) were standardized to 18S levels and expressed relative to M1 macrophages. Panel G: Immunostaining of eMPCs. Cells were stained with antibodies (red channel) to plasma membrane proteins differentially expressed by BmDCs (MBC2, FER1L), BmMs (ITGA6, STAB1), M2 cells (TFRC, ITGB5), and M1 cells (CD11a, CD40). Nuclei were visualized by DAPI staining (blue channel). Immunostaining and microscopy were performed on the same day and with identical microscope settings to experiments presented in Fig. 3D and Fig. S3 . Results obtained for flow cytometry, qRT-PCR, and immunocotyochemistry are representative of 3 independent analyses.

    Journal: PLoS ONE

    Article Title: Unique Proteomic Signatures Distinguish Macrophages and Dendritic Cells

    doi: 10.1371/journal.pone.0033297

    Figure Lengend Snippet: Mass spectrometric and immunohistochemical staining of thioglycolate-elicited peritoneal cells (eMPCs), polarized macrophages, and DCs. Panels A–B: Hierarchical cluster analysis of eMPCs. Cells were harvested from the peritoneal cavity of C57BL/6J- Ldlr tm1Her mice 5 days after intraperitoneal injection with thioglycolate. Isolated plasma membrane proteins detected by LC-MS/MS analysis of eMPCs were subjected to hierarchical cluster analysis, using the 107 proteins identified as differentially expressed by myeloid cells generated in vitro ( Fig. 2 ). Panel B: Relationships among eMPCs, M1 cells, M2 cells, BmMs, and DCs, as determined by cluster analysis. Panel C: Protein expression in eMPCs, M1 cells, M2 cells, BmMs, and BmDCs. Protein levels were quantified by MS/MS and spectral counting. Data are presented as means and SDs. Panels D–E: Flow cytometric analysis of CD11b, CD11c, and F4/80 in eMPCs. Results are presented as contour plots with 10% probability increments. Panel F: qRT-PCR analysis of M1 marker genes ( Nos2 , Il12b , Tnfa ) in M1 macrophages, BmDCs, and eMPCs. Results (means and SEMs; N = 6) were standardized to 18S levels and expressed relative to M1 macrophages. Panel G: Immunostaining of eMPCs. Cells were stained with antibodies (red channel) to plasma membrane proteins differentially expressed by BmDCs (MBC2, FER1L), BmMs (ITGA6, STAB1), M2 cells (TFRC, ITGB5), and M1 cells (CD11a, CD40). Nuclei were visualized by DAPI staining (blue channel). Immunostaining and microscopy were performed on the same day and with identical microscope settings to experiments presented in Fig. 3D and Fig. S3 . Results obtained for flow cytometry, qRT-PCR, and immunocotyochemistry are representative of 3 independent analyses.

    Article Snippet: Antibodies against murine CD11a (Abcam), CD11b (Abcam), CD11c (Abcam), CD40 (Abcam), MBC2 (Abcam), F4/80 (Abcam), ITGA6 (Abcam), ITGB5 (Abcam), MAC2 (Cedarlane), FER1L (Abcam), STAB1 (Santa Cruz), and TFRC (Abcam) were used in this study.

    Techniques: Immunohistochemistry, Staining, Mouse Assay, Injection, Isolation, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Generated, In Vitro, Expressing, Flow Cytometry, Quantitative RT-PCR, Marker, Immunostaining, Microscopy, Cytometry