Anti-CD206 antibody

• WB

Lab

Western blot - Anti-CD206 antibody (AB64693)

All lanes:

Western blot - Anti-CD206 antibody (ab64693) at 1 µg/mL

Lane 1:

Rat liver tissue lysate at 10 µg

Lane 2:

Human lung tissue lysate at 10 µg

Lane 3:

Rat lung tissue lysate at 10 µg

Lane 4:

Western blot - Mouse lung normal tissue lysate - total protein (<a href='/en-us/products/tissue-lysates/mouse-lung-normal-tissue-lysate-total-protein-ab29297'>ab29297</a>) at 10 µg

Secondary

All lanes:

Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/5000 dilution

Predicted band size: 166 kDa

Observed band size: 190 kDa

true

Exposure time: 4min

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD206 antibody (AB64693)

Immunohistochemical analysis of formalin fixed paraffin embedded human lung labelling CD206 with ab64693 at 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab64693 anti CD206 antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD206 antibody (AB64693)

ab64693 staining CD206 in MOLT4 cells. The cells were fixed with 4% Formaldehyde (10 min) permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab64693 at 1μg/ml and ab7291 Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488) pre-adsorbed at 1/1000 dilution (shown in green) and ab150120 Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594) pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD206 antibody (AB64693)

IHC image of ab64693 staining in human lung formalin fixed paraffin embedded tissue section*, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab64693, 0.1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD206 antibody (AB64693)

IHC image of CD206 staining in a section of formalin-fixed paraffin-embedded normal rat lung performed on a Leica BOND^TM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6 epitope retrieval solution 1) for 20mins. The section was then incubated with ab64693 0.01ug/ml for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions primary antibody concentration and antibody incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD206 antibody (AB64693)

IHC image of CD206 staining in a section of formalin-fixed paraffin-embedded normal mouse lung performed on a Leica BOND^TM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6 epitope retrieval solution 1) for 20mins. The section was then incubated with ab64693 0.01ug/ml for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions primary antibody concentration and antibody incubation times.

• IHC-P

AbReview66363****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD206 antibody (AB64693)

Paraffin embedded formalin-fixed mouse lung tissue labelling the CD206 with ab64693 at 1/500 in immunohistochemical analysis. An Alexa Fluor^® 647 Goat anti-rabbit IgG was used as the secondary antibody this was artificially colored green by software to enhance stain visibility. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. 15% serum was used as blocking agent for 1 hour at room temperature.

This image is courtesy of an anonymous Abreview

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-CD206 antibody (AB64693)

Immunocytochemistry/ Immunofluorescence using Anti-Mannose Receptor antibody, ab64693. Publication image from Zhao, S. J. et al., 2020, Theranostics, 31903103. Legend direct from paper.

Depletion of MSR1 in BMDMs reduced M2-like macrophages and mitochondrial OXPHOS in the co-culture system. (A) IF staining results of MSR1 WT and KO macrophages in the co-culture system for an M1-like marker (iNOS) and M2-like marker (CD206). Bar = 50 µm. (B) mRNA expression levels of M2-like marker genes (CD206 and CD163) in MSR1 WT and KO macrophages with or without co-culture by qPCR. Values are mean ± SD, *p < 0.05, ns indicates no significance. (C) Flow cytometry analysis of MSR1 WT or KO macrophages after co-culturing with BMSCs. Dot plots represent F4/80 and iNOS staining (left panel) and F4/80 and CD206 staining of macrophages (right panel). (D) Percentages of F4/80+ iNOS+ and F4/80+ CD206+ macrophages with or without co-culture were determined. Values are mean ± SD, ***p < 0.001, ns indicates no significance. (E) OCR of BMDMs in MSR1 WT or KO group after co-culture was detected using a Seahorse Bioscience XFp analyzer. O : Oligomycin, F : FCCP, A&R : antimycin A/rotenone. (F) Mitochondrial activities such as basal respiration, ATP production, respiratory capacity, and respiratory reserve were determined in indicated groups. Values are mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, ns indicates no significance.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-CD206 antibody (AB64693)

Immunocytochemistry/ Immunofluorescenceusing Anti-Mannose Receptor antibody, ab64693. Publication image from Zhao, S. J. et al., 2020, Theranostics, 31903103. Legend direct from paper.

MSR1-depletion reduced the fraction of M2-like macrophages on day 7 post-surgery in the tibial monocortical defect model. (A) Immunofluorescence (IF) staining of the total macrophage biomarker, F4/80 (green), and M1-like macrophage biomarker, iNOS (purple) and M2-like macrophage biomarker, CD206 (red), in facture tissues on day 3 post-surgery in the tibial monocortical defect model. Nuclei were counterstained with DAPI (blue). Bar = 200 µm. (B and C) The infiltration of F4/80+ macrophages (B) and the fraction of iNOS+ F4/80+ and CD206+ F4/80+ macrophages (C) were determined on day 3 post-surgery in the tibial monocortical defect model from MSR1 WT and KO mice (Values are expressed as mean ± SD, ns indicates no significance). iNOS group indicates the samples stained with anti-F4/80 and anti-iNOS; the slides stained with anti-F4/80, and anti-CD206 denote the CD206 group. (D) Representative IF images of total macrophages (F4/80+), M1-like macrophages (iNOS+ F4/80+), and M2-like macrophages (CD206+ F4/80+) in facture tissues on day 7 post-surgery of the tibial monocortical defect model. Nuclei were counterstained with DAPI (blue). Bar = 200 µm. (E) The iNOS+ and CD206+ macrophage fractions were determined by the percentages of iNOS+ and CD206+ macrophages within F4/80+ macrophage populations in the MSR1 WT or KO fracture tissues on day 7 post-surgery in the tibial monocortical defect model (Values are expressed as mean ± SD, *p < 0.05, **p < 0.01). (F) mRNA expression levels of macrophage marker genes (M1-like : iNOS and IL-1b, M2-like : CD206 and CD163) in the fracture tissues from MSR1 WT or MSR1 KO mice on day 7 in the tibial monocortical defect model (L) (*p < 0.05, **p < 0.01, ***p < 0.001).

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-CD206 antibody (AB64693)

Immunocytochemistry/ Immunofluorescence using Anti-Mannose Receptor antibody, ab64693. Publication image from Zhao, S. J. et al., 2020, Theranostics, 31903103. Legend direct from paper.

Overexpression of MSR1 in RAW264.7 cells promotes mitochondrial OXPHOS and M2-like polarization in the co-culture system. (A) OCR of blank (BL), vector (Vec), and MSR1-overexpressed (OE) RAW264.7 cells after co-culture were evaluated using a Seahorse Bioscience XFp analyzer. (B) Basal respiration, ATP production, respiratory capacity, and respiratory reserve were determined in BL, Vec, and MSR1 OE RAW264.7 cells with or without co-culture. Values are expressed as mean ± SD, **p < 0.01, ***p < 0.001, ns indicates no significance. (C) IF staining of BL, Vec, and OE RAW264.7 cells after co-culture for the M1-like biomarker (iNOS) and M2-like biomarker (CD206). Bar = 50 µm. (D) mRNA expression levels of M2-like macrophage marker genes (CD206 and CD163) in indicated groups were analyzed by qPCR. Values are expressed as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, ns indicates no significance. (E and G) Flow cytometry analysis of RAW264.7 cells from different groups after co-culture with BMSCs. Dot plots represent F4/80 and iNOS staining of (E) and F4/80 and CD206 staining of RAW264.7 cells (G). (F and H) percentages of F4/80+ iNOS+ (F) and F4/80+ CD206+ (H) in RAW264.7 cells in different groups were calculated. Values are expressed as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, ns indicates no significance.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-CD206 antibody (AB64693)

Immunocytochemistry/ Immunofluorescenceusing Anti-Mannose Receptor antibody, ab64693. Publication image from Zhao, S. J. et al., 2020, Theranostics, 31903103. Legend direct from paper.

Substitution of MSR1 KO bone marrow with MSR1 WT bone marrow promotes intramembranous bone healing. (A) Schematic representation of the main steps of the bone marrow transplant. (B) Representation of 3D images of injured tibiae from different transplanted mice (KO to KO vs. WT to KO) by micro-CT on day 7 or 14 post-surgery. (C-F) Quantification of BV/TV (%) (C), Tb. Th (mm) (D), Tb. N (/mm) (E) and Tb. Sp (mm) (F) in the defect region on day 7 and 14 post-surgery for different transplanted mice (KO to KO vs. WT to KO) (Values are expressed as mean ± SD, *p < 0.05). (G) Representative IF images of total macrophages (F4/80+), M1-like macrophages (iNOS+ F4/80+) and M2-like macrophages (CD206+ F4/80+) in facture tissues from the bone marrow of transplanted mice (transplanted MSR1 KO bone marrow to host KO mice and transplanted MSR1 WT bone marrow to host KO mice) on day 7 post-surgery in the tibial monocortical defect model. Nuclei were counterstained with DAPI (blue). Bar = 100 µm. (H) The iNOS+ and CD206+ macrophage fractions were determined from different transplanted mice (KO to KO vs. WT to KO) on day 7 post-surgery of the tibial monocortical defect model (Values are expressed as mean ± SD, *p < 0.05, **p < 0.01). (I) mRNA expression levels of macrophage marker genes (M1-like : iNOS and IL-1b, M2-like : CD206 and CD163) in fracture tissues from different transplanted mice on day 7 in the tibial monocortical defect model (K) (*p < 0.05, **p < 0.01, ***p < 0.001).

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-CD206 antibody (AB64693)

Immunocytochemistry/ Immunofluorescence using Anti-Mannose Receptor antibody, ab64693. Publication image from Zhao, S. J. et al., 2020, Theranostics, 31903103. Legend direct from paper.

MSR1-depletion reduced the fraction of M2-like macrophages on day 7 post-surgery in the tibial monocortical defect model. (A) Immunofluorescence (IF) staining of the total macrophage biomarker, F4/80 (green), and M1-like macrophage biomarker, iNOS (purple) and M2-like macrophage biomarker, CD206 (red), in facture tissues on day 3 post-surgery in the tibial monocortical defect model. Nuclei were counterstained with DAPI (blue). Bar = 200 µm. (B and C) The infiltration of F4/80+ macrophages (B) and the fraction of iNOS+ F4/80+ and CD206+ F4/80+ macrophages (C) were determined on day 3 post-surgery in the tibial monocortical defect model from MSR1 WT and KO mice (Values are expressed as mean ± SD, ns indicates no significance). iNOS group indicates the samples stained with anti-F4/80 and anti-iNOS; the slides stained with anti-F4/80, and anti-CD206 denote the CD206 group. (D) Representative IF images of total macrophages (F4/80+), M1-like macrophages (iNOS+ F4/80+), and M2-like macrophages (CD206+ F4/80+) in facture tissues on day 7 post-surgery of the tibial monocortical defect model. Nuclei were counterstained with DAPI (blue). Bar = 200 µm. (E) The iNOS+ and CD206+ macrophage fractions were determined by the percentages of iNOS+ and CD206+ macrophages within F4/80+ macrophage populations in the MSR1 WT or KO fracture tissues on day 7 post-surgery in the tibial monocortical defect model (Values are expressed as mean ± SD, *p < 0.05, **p < 0.01). (F) mRNA expression levels of macrophage marker genes (M1-like : iNOS and IL-1b, M2-like : CD206 and CD163) in the fracture tissues from MSR1 WT or MSR1 KO mice on day 7 in the tibial monocortical defect model (L) (*p < 0.05, **p < 0.01, ***p < 0.001).