Three slices were randomly selected from Sham group and SAH group of ISF tracking respectively, and the immunofluorescence staining was performed as reported by others [7 (link)]. Briefly, brain sections including frontal cortex were incubated with anti-GFAP (1:300, ab7260, Abcam, Cambridge, UK), anti-α-SMA (1:200, A5228, Sigma-Aldrich, Darmstadt, Germany), anti-MMP9 (1:200, ab76003, Abcam, Cambridge, UK), anti-COL4A3 (1:200, ABIN678128, Antibodies-online, Aachen, Germany), and corresponding secondary antibodies (Alexa Fluor 488 goat anti-mouse, 1:500, #4408, Cell Signaling Technology, Massachusetts, USA; Alexa Fluor 488 goat anti-rabbit, 1:500, #4412, Cell Signaling Technology, Massachusetts, USA). The images of the staining were captured using a Laser Scanning Confocal Microscope (LSCM) SP8 (Leica, Heerbrugg, Germany).
Alexa fluor 488 goat anti mouse
Manufactured by Cell Signaling Technology
Sourced in United States
Alexa Fluor 488 goat anti-mouse is a secondary antibody conjugated with the Alexa Fluor 488 fluorescent dye. It is designed to detect and bind to mouse primary antibodies, allowing for visualization and detection of target proteins or molecules in various biological applications.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 goat anti rabbit, by Cell Signaling Technology (2 mentions)
Alexa fluor 594 goat anti mouse, by Cell Signaling Technology (2 mentions)
Alexa fluor 594 goat anti rabbit, by Cell Signaling Technology (2 mentions)
Lsm 880, by Zeiss (2 mentions)
Ab76003, by Abcam (1 mentions)
Ab7260, by Abcam (1 mentions)
A5228, by Merck Group (1 mentions)
Cyclin b1, by Cell Signaling Technology (1 mentions)
Ab22609, by Abcam (1 mentions)
Anti c fos, by Abcam (1 mentions)
Mab377, by Abcam (1 mentions)
Anti flag, by GenScript (1 mentions)
Anti ha 3724, by Cell Signaling Technology (1 mentions)
A11012, by Cell Signaling Technology (1 mentions)
Anti cd16 apc, by BioLegend (1 mentions)
Rpmi 1640 medium, by Sartorius (1 mentions)
Anti cd11b pe, by BioLegend (1 mentions)
Alexa fluor 568 goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Sartorius (1 mentions)
Anti cd66b fitc, by BioLegend (1 mentions)
6 protocols using alexa fluor 488 goat anti mouse
1
Immunofluorescence Analysis of Brain Sections
2
Immunofluorescence Analysis of DNA Damage Response in Embryonic Mouse Tissues
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Embryos of pregnant female mice at E11.5 and E12.5 were used for the study. Embryonic mouse tissues were fixed in 4% paraformaldehyde and paraffin-embedded to perform continuous tissue sectioning at a thickness of 4 μm. Tissue sections were dewaxed, dehydrated, washed in PBS, and then subjected to antigen repair in Tris-EDTA (pH = 8.0) solution at high temperature and pressure for 15 min. The samples were incubated with 10% goat serum containing 0.3% TritonX-100 for 1 h at room temperature to permeabilize and block the samples. Tissue sections were incubated with primary antibody dilution overnight at 4℃, washed three times with PBS, and incubated with secondary antibody dilution for 1 h at room temperature. After washing with PBS, the sections were closed with anti-quenching reagent containing DAPI and observed under fluorescence microscopy. Antibodies used in the study included: SSEA1 (ab16285), PARP3 (PA5-112641), NEIL2 (PA5-103829), BLM (PA5-27384), RAD51 (NB100-148), LIG1 (MA5-42920), PCNA (SC-56), DNA-RNA Hybrid [S9.6] (Kf-Ab01137-23.0), FANCD2 (NB100-182), Cyclin B1(Cell Signaling, 4138), Alexa Fluor®594 Goat Anti-Rabbit (ab150080), Alexa Fluor®488 Goat Anti-Mouse (ab150117), Alexa Fluor®594 Goat Anti- Mouse (ab150116).
3
Immunofluorescence Staining of Brain Sections
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Tissue sections were incubated in 0.3% Triton X-100 for 20 min and then blocked with 10% goat serum for 45 min at room temperature (20° to 23°C). The sections were incubated with primary antibodies, including anti–c-Fos (1:1000; Abcam, ab190289), anti-MOR (1:300; EMD Millipore, AB5509), anti-CCK2R (1:500; Alomone Labs, ACR-042), anti-CCK8s (1:300; IMMUNOSTAR, 20078), anti-NeuN (1:1000; Sigma-Aldrich, MAB377), and anti-CaMKII (1:250; Abcam, ab22609) at 4°C for 24 hours. After the incubation with primary antibodies, sections were washed with PBS (5 × 8 min) and then incubated with corresponding fluorophore-conjugated secondary antibodies, including Alexa Fluor 594 goat anti-rabbit (1:500; Cell Signaling Technology, 8889S), Alexa Fluor 594 goat anti-pig (1:300; Invitrogen, A11075), Alexa Fluor 488 goat anti-mouse (1:500; Cell Signaling Technology, 4880S), Alexa Fluor 488 goat anti-rabbit (1:500; Cell Signaling Technology, 4412S), and Alexa Fluor 594 goat anti-mouse (1:500; Cell Signaling Technology, 8890S) for 2 hours at room temperature. For c-Fos immunostaining, HC-mice and CC-mice were anesthetized in the HC and CC 90 min after the last morphine injection on day 5, respectively, followed by transcardially perfusion. Brian slices were examined and photographed using an LSM 880 (Zeiss, Germany) confocal microscope.
4
Colocalization of CD2v and CD58 in Cells
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PK15 cells were cotransfected with pCD2v-HA and swine pCD58-Flag, fixed with 4% PFA 24 h pt, permeabilized with 0.2% Tritron-X 100 and treated with anti-HA (3724S; Cell Signaling Technology, MA, USA) or anti-Flag (A00187; Genscript, Piscataway, NJ, USA) primary antibodies. Cells were then washed with PBS and incubated with goat anti-rabbit Alexa fluor 594 (A11012; Cell signaling Technology, Danvers, MA, USA) and goat anti-mouse Alexa fluor 488 (A11029; Cell Signaling Technology, Danvers, MA, USA) secondary antibodies for 1 h. Nuclei were stained with DAPI and images obtained using the A1 Nikon confocal microscope. Colocalization of CD2v with endogenous human CD58 was investigated in 293T cells. Cells were transfected with pCD2v-HA and IFA was performed using anti-HA (3724S; Cell Signaling Technology, Danvers, MA, USA) and anti-CD58 (sc20009; Santa Cruz, CA, USA) primary antibodies, and goat anti-rabbit Alexa fluor 594 (A11012; Cell Signaling Technology, MA, USA) and goat anti-mouse Alexa fluor 488 (A11029) secondary antibodies.
5
Neutrophil Activation and Apoptosis Assays
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Fetal bovine serum (FBS), goat sera, RPMI-1640 medium and PBS were purchased from Biological Industries Israel Beit Haemek. X-VIVO™ 15 medium was purchased from Lonza (cat. no. 04-418Q). FITC Annexin V Apoptosis Detection kits were purchased from BD Pharmingen (cat. no. 556547). Anti-CD11b-PE, anti-CD16-APC and anti-CD66b-FITC were purchased from Biolegend (cat. no. 101207, 302011 and 305104, respectively). ATRA was purchased from MedChemExpress (cat. no. 302-79-4). DMSO was purchased from Sigma-Aldrich (Merck KGaA; cat. no. D8418-100). Neutrophil isolation kits were purchased from TBD (cat. no. LZS11131). citH3 antibody (histone H3 citrulline R2 + R8 + R7), anti-MPO antibody and Ca2+ ionophore (CI) were purchased from Abcam (cat. nos. ab5103, ab9535 and ab120287, respectively). Goat anti-mouse Alexa Fluor 488, ProLong Gold Antifade reagent and histone H3 mouse antibody were purchased from Cell Signaling Technology, Inc. (cat. nos. 4408s, 9071S and 14269s, respectively). Goat anti-rabbit Alexa Fluor 568, Total ROS Assay kits and Quant-iT™ PicoGreen™ double-strand (ds)DNA Assay kits were purchased from Thermo Fisher Scientific, Inc. (cat. no. A-11011, 88-5930-74 and P7589, respectively). R-PE-antibody conjugation kits were purchased from Fcmacs (cat. no. FMS-ABPE0001).
6
Quantification of IGF-II/IGF2R Expression
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A total of 2.5 × 105 Sol8 cells (WT, bulk populations, E2_1 and transduced E2_1 cells) were plated in 6-well plates with 2 mL of DMEM (supplemented with 20% FBS and 1% penicillin/streptomycin). After 24 h, the cells were detached using cell scrapers and kept on ice (4 °C) to avoid internalisation of CI-MPRs. A total of 105 cells were washed twice with cold PBS + 1% BSA and resuspended in blocking buffer (PBS + 1% BSA with goat serum) for 30 min on ice. After two washes, cells were stained with anti IGF-II/IGF2R mouse antibody (2G11) (#NB300-514, Novus biologicals, Centennial, CO, USA) for 1.5 h on ice at 4 °C. Mouse IgG2a kappa was used as an isotype control. Cells were washed twice and incubated with a secondary antibody goat anti-mouse AlexaFluor-488 (#4408S, Cell Signaling Technology, Danvers, MA, USA) for 1 h on ice (dark). Cells were washed two times and analysed on a FACS Canto II flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA) using the FACS Diva software (BD Biosciences, Bedford, MA, USA) and FlowJo program (BD, Ashland, OR, USA).
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