To reduce nonspecific binding to cells bearing Fc receptors, preincubation of cells with anti-mouse CD16/CD32 (eBioscience) was performed prior to all antibody stainings. Dead cells were excluded by FxCycle Violet (Thermo Fisher Scientific) staining. Stained cells were analyzed using a BD FACSCanto II or analyzed and FACS-sorted using a BD Aria cell sorting platform. The frequencies of individual cell populations were quantified with FlowJo software. FACS antibodies used in this study were: rat anti-CD11b (eBioscience, PE-Cy7), rat anti-CD31 (BioLegend, FITC), rat anti-F4/80 (BioLegend, PE), rat anti-Ly6C (BioLegend, APC-Cy7), rat anti-Ly6G (BioLegend, Pacific Blue), rat anti-Tie2 (Invitrogen, APC) and rat anti-CD206 (BD Biosciences, Alexa Fluor 488). Single-stained controls and an unstained control were used for compensation, rat-IgG1 κ (eBioscience, APC) was used as isotype control.
Rat anti cd11b
Manufactured by Thermo Fisher Scientific
Sourced in United States
Rat anti-CD11b is a laboratory reagent used for the identification and analysis of cells expressing the CD11b antigen. CD11b is a cell surface marker expressed on various immune cells, including monocytes, macrophages, and granulocytes. This antibody can be used in techniques such as flow cytometry to detect and quantify CD11b-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Rat anti cd45, by Thermo Fisher Scientific (3 mentions)
Abc kit, by Vector Laboratories (2 mentions)
Dab kit, by Vector Laboratories (2 mentions)
Tsa plus kit, by PerkinElmer (2 mentions)
M o m kit, by Vector Laboratories (2 mentions)
Alexa fluor 350 phalloidin, by Thermo Fisher Scientific (2 mentions)
Mouse anti β catenin, by BD (2 mentions)
Goat anti pcadherin, by R&D Systems (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Lsm 800, by Zeiss (2 mentions)
Fluorescent labeled secondary antibody, by Thermo Fisher Scientific (2 mentions)
Rabbit anti neun, by Abcam (2 mentions)
Chicken anti map2, by Abcam (2 mentions)
Rat igg1 κ, by Thermo Fisher Scientific (1 mentions)
Rat anti cd31, by BioLegend (1 mentions)
Rat anti f4 80, by BioLegend (1 mentions)
Facscanto 2, by BD (1 mentions)
Anti mouse cd16 cd32, by Thermo Fisher Scientific (1 mentions)
Fxcycle violet, by Thermo Fisher Scientific (1 mentions)
Rat anti ly6g, by BioLegend (1 mentions)
6 protocols using rat anti cd11b
1
Multiparametric Immunophenotyping of Myeloid Cells
2
Immunohistochemical Analysis of Skin Samples
3
Immunohistochemical Analysis of Skin Samples
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Skin was fixed in 4% PFA for whole mount or in 10% formalin for paraffin embedding and used for histological analysis as previously described26 . Immunohistochemistry was performed by incubating sections at 4°C overnight with primary antibodies as follows: mouse anti-β-catenin (1:100, BD #610153; 14/Beta-Catenin), rat anti-CD11b (1:250, eBioscience #14-0112; M1/70), goat anti-P-cadherin (1:100, R&D #AF761), rabbit anti-pSmad2 (Ser465/467) (1:1000, Cell Signaling #3108; 138D4), and rabbit anti-Lef-1 (1:100, Cell Signaling #2286; C18A7). pSmad2 immunostaining required TSA Plus kit (PerkinElmer). For brightfield immunohistochemistry, biotinylated species-specific secondary antibodies, followed by detection using the ABC kit (Vector Labs) and DAB kit (Vector Labs), were used according to the manufacturer’s instructions. M.O.M. kit was used for mouse antibodies (Vector Laboratories). Secondary antibodies conjugated with FITC, RRX and Cy5 (Jackson Immunoresearch Laboratories) were used at a concentration of 1:100 for 1 hour at room temperature. Alexafluor 350 phalloidin (Life Technologies) was used according to the manufacturer’s instructions.
4
Immunohistochemical Staining of Neural Markers
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Sections were washed 3 times with 1× PBS and then incubated with the primary antibodies at 4 °C overnight after 1 h blocking by 5% normal goat serum (NGS, Sigma) and 0.2% Triton X-100 (Sigma). The sections were then incubated at room temperature for 2 h with fluorescent-labeled secondary antibodies (Invitrogen, Waltham, MA, USA) and washed with 0.01 M PBS 3 times before being observed under a confocal laser scanning microscope (Zeiss, LSM800). Fluorescence immunohistochemistry was performed using the following primary antibodies: rabbit anti-NeuN (Abcam, Cambridge, MA, USA; 1:500), chicken anti-microtubule-associated protein 2 (Map2) (Abcam, 1:500), rabbit anti-glial fibrillary acidic protein (Gfap) (Dako, St. Clara, CA, USA; 1:1,000), rat anti-Cd45 (eBioscience, Waltham, MA, USA; 1:500), purified anti-Neurofilament Marker (pan axonal, cocktail, SMI-312) (Biolegend, San Diego, CA, USA; 1:1,000), mouse anti-fibronectin (Fn1) (Abcam, 1:200), rat anti-CD11b (eBioscience, 1:500), guinea pig anti-ionized calcium-binding adapter molecule 1 (IBA1) (Synaptic Systems, Goettingen, Germany; 1:800), rabbit anti-CD34 (Abcam, 1:250).
5
Spinal Cord Histology and Immunohistochemistry
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Animals were euthanized by overdose of pentobarbital. After transcardial perfusion with PBS and 4% paraformaldehyde, spinal cords were excised under a dissection microscope and post-fixed in paraformaldehyde at 4 °C for another 8 h, followed by cryoprotection using 30% sucrose. The spinal cord tissue including lesion area was embedded in OCT compound and sliced horizontally to produce 15 µm frozen sections using a cryostat microtome (Leica). Fluorescence immunohistochemistry was performed using following primary antibodies: rabbit anti-glia fibrillary acid protein (GFAP, 1:1,000, Dako), rat anti-CD45 (ebioscience, 1:1,000), rat anti-CD11b (ebioscience, 1:500), rabbit anti-IBA1 (WAKO, 1:500), rabbit anti-SNAP25 (Synaptic Systems GmbH, 1:1,000), purified anti-Neurofilament Marker (pan axonal, cocktail, SMI-312) (Biolegend, 1:1,000), mouse anti-CNPase (Abcam, 1:1,000), rabbit anti-PLP1 (Abcam, 1:1,000), fluorescence labeled Lectin (Vector, 1:20), chicken anti-MAP2 (Abcam, 1:1,000), rat anti-NeuN (Abcam, 1:1,000).
6
Multimarker Immunohistochemistry Protocol
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Sections were washed three times with 1× PBS and then incubated with the primary antibodies at 4 °C overnight after 1 h blocking by 5% normal goat serum (NGS, Sigma) and 0.2% Triton X-100 (Sigma). The sections were then incubated at room temperature for 2 h with fluorescent-labeled secondary antibodies (Invitrogen) and washed with 0.01 M PBS 3 times before being observed under a confocal laser scanning microscope (Zeiss, LSM800). Fluorescence immunohistochemistry was performed using the following primary antibodies: rabbit anti-NeuN (Abcam, 1:500), chicken anti-MAP2 (Abcam, 1:500), rabbit anti-GFAP (Dako, 1:1000), chicken anti-GFAP (Abcam, 1:1000), rat anti-CD45 (ebioscience, 1:500), rat anti-CD11b (ebioscience, 1:500), guineapig anti-IBA1 (Synaptic Systems, 1:800), rabbit anti-SLC1A3 (EAAT1/GLAST) (Abcam, 1:500), mouse anti-AQP4 (Abcam, 1:350), rabbit anti-SOX2 (Abcam, 1:500), rabbit anti-MKI67 (Abcam, 1:500).
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