MA1-91124

Product number: MA1-91124 - Provider product page
Provider: Invitrogen Antibodies
Product name: F4/80 Monoclonal Antibody (CI:A3-1)
Antibody type: Monoclonal
Antigen: Other
Description: Positive control of mouse macrophages suggested. MA1-91124 detects a band of approximately 160 kDa due to the protein being heavily glycosylated, predicted molecular weight of precursor protein is 102 kDa. Store product as a concentrated solution. Centrifuge briefly prior to opening the vial. This antibody recognizes murine F4/80 antigen which shares 68% overall amino acid identity with human EGF module-containing mucin-like hormone receptor 1 (EMR1).
Reactivity: Human, Mouse
Host: Rat
Isotype: IgG
Antibody clone number: CI:A3-1
Vial size: 250 µg
Concentration: 1 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

ADGRE1 protein structure - MA1-91124 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7 FA injection induces interstitial cell infiltration in transgenic line A homozygous mice. The inflammatory cell infiltration was characterized by immunohistochemistry with anti-F4/80 (monocytes/macrophages) and anti-CD3 (T cells) antibodies. (A and B). Representative immunostaining of one mouse from each group (x400 magnification). (C) Quantification of positive IHC signals were quantified for (a) F4/80, (b) CD3 and (c) PCNA using KS300 image analyzer software. All parameters were significantly increased in transgenic mice. Data are shown as the mean +- SEM of 14-18 mice per group * p < 0.05; ** p < 0.01 vs WT-FA.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 miR-23a is upregulated in DRG neurons derived from mice after SNI. ( A ) miR-23a expression in contralateral L5 DRG neurons at 7 day after model establishment, normalized to U6 (Scale bar = 50 mum) and quantitative histogram of miR-23a + neurons using FISH; ( B ) miR-23a expression in DRG neurons of sham-operated and SNI mice were determined using RT-qPCR, normalized to U6; ( C ) macrophages in DRG (F4/80+ cells, red) and miR-23a (green) detected using FISH and immunohistochemistry (Scale bar = 50 mum); ( D ) miR-23a expression in DRG neurons treated with 0.001% DMSO in HEPES buffer + glucose (1 mg/mL) and with capsaicin (1 muM), normalized to U6. Values obtained from three independent experiments are expressed as mean +- SD and analyzed by unpaired t -test. * indicates p < 0.05. Cell experiment was independently repeated for three times.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 Delivery of decreased miR-23a by DRG neuron-derived EVs reduces neuropathic hypersensitivity and recruitment of M1 macrophages in vivo . SNI mice were treated with intrathecal administration of EVs-treated miR-23a antagomir-transfected DRG neurons. ( A ) effect of intrathecal injection of EVs-miR-23a antagomir on mechanical hypersensitivity for 7 days, which was expressed as mean +- SD of 50% PWT; n (vehicle mice) = 6, n (oligomer-treated mice) = 12; * p < 0.05 vs. vehicle mice; # p < 0.05 vs. oligomer-treated mice. Values obtained from three independent experiments in triplicate are expressed as mean +- SD and analyzed by two-way ANOVA, followed by Tukey''s test among multiple groups. ( B ) miR-23a expression in DRG determined using RT-qPCR, normalized to U6; ( C ) Immunostaining of macrophages (F4/80+ cells, red), and nuclei (DAPI, blue) in SNI DRG (scale bar = 25 mum); ( D ) Macrophage phenotyping [M1 macrophages (CD206 - CD11c + ) and M2 macrophages (CD206 + CD11c - )] in DRG analyzed using flow cytometry; ( E ) levels of TNF-alpha and IL-6 in L4-L5 DRG tissues measured using ELISA. Values obtained from three independent experiments are expressed as mean +- SD and analyzed by unpaired t -test between two groups in panel ( B - E ). * p < 0.05 vs. mice treated with EVs-miR-23a-NC or vehicle. Cell experiment was independently repeated for three times.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 The effects of MTL-CEBPA on MCD-induced fatty liver disease. a Histological analysis of liver sections from normal diet and MCD diet-fed animals for ( a , upper panels) 4 weeks and ( a , lower panels) 6 weeks stained with H&E and Masson's trichome (magnification: x 20). ( a , left panel) Analysis of liver sections from normal diet control animals showed normal architecture with hepatocytes arranged in the form of cords (blue arrow) radiating away from the central vein. The portal trip (red arrow) contained an artery, vein, and the bile duct. Normal distribution of the sinusoidal spaces (yellow arrow) was observed. ( a , right panel) Analysis of the liver sections from MCD diet animals revealed steatosis (green arrow) and inflammatory cell infiltration (black arrow). b Serum levels of ALT and AST indicate reduction to near normal levels following 2 weeks of MTL-CEBPA treatment. Bilirubin levels do not continue to increase in the presence of 1 and 3 mg/kg of MTL-CEBPA treatment. Y= = p (0.0002); * = p (0.002); $ = p (0.003). Histological analysis of liver sections from MCD diet-fed animals treated with NOV340/FLUC (3 mg/kg) and MTL-CEBPA at 0.3 mg/kg; 1.0 mg/kg and 3 mg/kg. c Liver sections stained with H&E and Masson's trichrome. (magnification: x 20). Analysis of liver sections from MCD + NOV340/FLUC revealed steatosis (green arrow) and inflammatory cell infiltration (black arrow) which was more prominent, when compared with MCD diet at week 6. The liver sections from

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIG 5 Rapid kinetics of liver Kupffer cell depletion followed by low levels of inflammatory cell infiltration to liver in mice following intravenous injection of GAd. Immunofluorescence microscopy analysis of F4/80 + macrophages (top two rows) and CD8 + T cells (bottom two rows) in liver from untreated C57BL/6J mice (untreated, n = 3) and from mice at days 1, 3, 5, 7, and 11 following intravenous injection of 1.0 x 10 11 viral particles (vp) of HAd5 and GAd ( n = 2 for each virus and time point). Magnifications, x100. Red, CD31/endomucin; green, GFP; blue, DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIG 7 Lack of substantial inflammatory cell infiltration into lung in mice following intravenous injection of GAd. Immunofluorescence microscopy was used to analyze F4/80 + macrophages and CD8 + T cells in lungs from untreated C57BL/6J mice (untreated, n = 3) and from mice at days 1, 3, 7, and 11 following intravenous injection of 1.0 x 10 11 viral particles (vp) of HAd5 and GAd ( n = 2 for each virus and time point). Magnifications, x100. Red, CD31/endomucin; green, GFP; blue, DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 5 FIGURE In vivo: AMD3100 inhibited LPS-stimulated M1 macrophage polarization. (A, B) Fluorescence confocal microscopy analysis of M1 macrophage polarization. Representative images of IF showing DAPI, F4/80 +ve , iNOS +ve and merged images. Nuclei were stained by DAPI and are shown in blue. F4/80 marks macrophages while iNOS is a marker of M1 macrophages. (A) Uterus tissue from mice treated with PBS, LPS or LPS + AMD3100 respectively. (B) Macrophages treated with PBS, LPS, CXCL12, LPS-SMC-CM and LPS-SMC-CM + AMD3100 respectively. (C) Quantitative analyses of macrophage polarization (iNOS +ve cells) in uterus showing significant increase in polarization in LPS-treated macrophages compared to PBS. Treatment with AMD3100 inhibited the polarization stimulated by LPS. (D) Quantitative analyses of macrophage polarization (iNOS +ve cells) in vitro showing significant increase in polarization in LPS, CXCL12 and LPS-SMC-CM treated macrophages compared with PBS treatment. AMD3100 inhibited the polarization stimulated by LPS. Results shown as mean +- SE. * p < 0.05 versus PBS while # p < 0.05 versus LPS-SMC-CM. Scale bar: 75 mum. DAPI, 4,6-diamidino-2-phenylindole; IF, immunofluorescence; IHC, immunohistochemistry; LPS, lipopolysaccharide; PBS, phosphate buffered saline; SE, standard error; SMC-CM, smooth muscle cell conditioned media

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Increased M2Mph by co-culture with hAM-MSCs in vitro Mouse bone marrow-derived Mph were co-cultured with hAM-MSCs using a Transwell insert (0.4 mum pore size) for 48 h to investigate the role of hAM-MSC secretome on Mph activation. (A) Representative flow cytometry dot plots of Mph with or without co-culture with hAM-MSCs (Mph + hMSC and Mph groups, respectively). Cells double positive for F4/80 and CD206 within the CD11b + population were defined as M2Mph. Percentage of M2Mph among the total live cells as well as the absolute number of M2Mph was calculated and presented in the charts. (B) Immunofluorescent images for F4/80, CD206, and DAPI of Mph, demonstrating enhanced CD206 expression by co-culture with hAM-MSCs compared to the ordinary Mph monoculture. Scale bars, 20 mum. (C) RT-PCR quantification of Mph gene expression after hAM-MSC co-culture (Mph + hMSC group) compared to the Mph-only group using GAPDH as an internal reference gene. Fold change in mRNA levels was normalized to 1 for the control group. n = 4 biological replicates with 3 technical replicates. (D) Secretion levels of IL-10 and TGF-beta1 of Mph with or without co-culture with hAM-MSCs. n = 4 biological replicates with 3 technical replicates. Data are shown as mean +- SEM. *p < 0.05, **p < 0.01, and ***p < 0.001. Student's t test (A and C); one-way ANOVA with Tukey's post hoc tests (D).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2. Immune cell profiles in axotomized DRGs. ( A ) Representative images of L4 DRG cross sections from naive mice, d1, d3, and d7 post-SNC. Macrophages were stained with anti-Iba1 and anti-F4/80. Neurons were stained with anti-NFH. Scale bar, 50 um. ( B ) Quantification of granulocytes per DRG detected by flow cytometry. For flow cytometry of DRGs, naive mice (n = 14 biological replicates), d1 (n = 3), d3 (n = 5), and d7 (n = 12) post-SNC mice were used. Granulocytes (CD45 + , CD11b + , Ly6G + ) per DRG are shown. ( C ) Quantification of Mo/Mac (CD45 + , CD11b + , CD11c - , Ly6G - ) per DRG. ( D ) Quantification of MoDC (CD45 + , CD11b + , CD11c + , Ly6G - ) per DRG. ( E ) Quantification of cDC (CD45 + , CD11b - , CD11c + , Ly6G - ) per DRG. ( F ) Quantification of lymphocytes (CD45+, CD11b - ) per DRG. ( G ) Composition of CD45 + leukocytes in lumbar DRGs identified by flow cytometry. Flow data are represented as mean cell number +- SEM. Each data point represents L3-L5 DRGs pooled from three to four animals (18-24 DRGs), biological replicates, n = 3-14. Statistical analysis was performed in GraphPad Prism (v7) using one-way or two-way ANOVA with correction for multiple comparisons with Tukey's post-hoc test. For B-F, unpaired two-tailed t-test with Welch's correction. A p value < 0.05 (*) was considered significant. p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3. Sciatic nerve injury triggers massive accumulation of hematogenous leukocytes in the injured nerve but not axotomized DRGs. ( A ) Parbiosis complex of a wildtype (WT) and a tdTomato (tdTom) mouse. Mice were surgically paired at postnatal day 56. The timeline of the experiment is shown. ( B ) Flow cytometric analysis of sciatic nerve trunks collected from non-parabiotic (single) tdTom mice, WT parabionts, and tdTom parabionts. Dotplot of live (CD11b + , tdTom + ) cells in the d3 post-SNC nerve. ( C ) Quantification of tdTom + myeloid cells in the 3d injured nerve of WT single mice (WT-S), WT parabiont (WT-para), tdTom parabiont (tdTom-para), and tdTom single (tdTom-S) mice. The fraction of tdTom + myeloid cells (CD45 + , CD11b + ), MoDC (CD45 + , CD11b + , CD11c + , Ly6G - ), and Mo/Mac (CD45 + , CD11b + , CD11c - , Ly6G - ) is shown. For quantification of tdTom + immune cells, nerves from the WT parabiont and the tdTom parabiont were harvested separately (three mice per data point) with n = 2-3 biological replicates. Flow data are represented as fraction of tdTom + cells +- SEM. Statistical analysis was performed in GraphPad Prism (v8) using one-way ANOVA with correction for multiple comparisons with Tukey's post-hoc test. p value of < 0.001 (***) and p