11-mercaptoundecanoic acid (MUA), 2-(N-morpholino) ethanesulfonic acid (MES), serum-coloring agents, protein agents, trifluoroacetic acid, hexafluoro-2-propanol (HFIP), rabbit anti-mouse IgG/FITC, and bovine serum albumin (BSA) were obtained from Sigma-Aldrich. Hydroxysuccinimide (NHS) and 1-ethyl-3-(3-dimethylami-nopropyl) carbodiimide hydrochloride (EDC) were purchased from Acros-Organics. K3[Fe(CN)6]·3H2O and K4[Fe(CN)6] were obtained from SHOWA Inc. 10X Phosphate buffered saline (PBS) buffer was purchased from GeneMark Inc. LC5800 pre-stained protein, Aβ(1–40) antigens, and Aβ(1–42) antigens were purchased from ENZO Life Science. Aβ(1–42) monoclonal antibodies (12F4), monoclonal antibodies (4G8), and immunoglobulin G (IgG) were obtained from NOVUS Inc. All chemicals were used without further purification.
Rabbit anti mouse igg fitc
Manufactured by Merck Group
Rabbit anti-mouse IgG FITC is a laboratory reagent that contains antibodies specific to mouse immunoglobulin G (IgG) molecules. These antibodies are conjugated with the fluorescent dye fluorescein isothiocyanate (FITC), allowing for the detection and visualization of mouse IgG in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Sheep anti mouse c3 fitc, by Merck Group (2 mentions)
Accuri, by BD (2 mentions)
Rabbit anti human cd133 prominin 1 antibody, by Merck Group (2 mentions)
Mouse anti human cd44 conjugated with fitc, by Thermo Fisher Scientific (2 mentions)
Mouse anti human cd24 antibody conjugated with rpe, by Thermo Fisher Scientific (2 mentions)
1 ethyl 3 3 dimethylaminopropyl carbodiimide hydrochloride edc, by Thermo Fisher Scientific (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
11 mercaptoundecanoic acid, by Merck Group (1 mentions)
Hexafluoro 2 propanol hfip, by Merck Group (1 mentions)
Percoll gradient, by Merck Group (1 mentions)
Collagenase 4, by Merck Group (1 mentions)
Dnase 1, by Roche (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Cell strainer, by BD (1 mentions)
5 protocols using rabbit anti mouse igg fitc
1
Amyloid-Beta Protein Detection Assay
2
Murine Lupus Nephritis Evaluation
3
Spleen and Kidney Tissue Analysis
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Spleens were harvested and kept on ice during processing. The spleens were processed through 40 μm strainers and depleted of red blood cells with red blood cell lysis buffer (144 mM NH4Cl and 17 mM Tris, pH 7.6). Spleen cells were washed twice with cold RPMI before being stained for flow cytometry analysis.
One kidney was divided evenly for renal pathology and immunofluorescence. One half was snap frozen in liquid nitrogen and stored at −80°C for immunofluorescence analysis, the other half was fixed with buffered formalin, embedded in paraffin, and then sectioned and stained with hematoxylin and eosin. Kidney sections were analyzed in a blinded fashion by Dr. Phillip Ruiz (Department of Pathology, University of Miami School of Medicine, Miami, FL, USA) and graded on glomerular hypercellularity, segmental mesangial expansion, neutrophils/cell debris, crescent formation, and necrosis. These grades were combined for a total glomerular and interstitial pathology score. Deposition of IgG and complement component C3 was analyzed on the half kidney snap frozen in liquid nitrogen. Deposition was assessed by immunofluorescence by incubating slides with rabbit anti-mouse IgG FITC (Sigma) and sheep anti-mouse C3 FITC (Sigma). IgG and C3 were graded 0–3 for intensity of staining, as previously described (4 (link)).
One kidney was divided evenly for renal pathology and immunofluorescence. One half was snap frozen in liquid nitrogen and stored at −80°C for immunofluorescence analysis, the other half was fixed with buffered formalin, embedded in paraffin, and then sectioned and stained with hematoxylin and eosin. Kidney sections were analyzed in a blinded fashion by Dr. Phillip Ruiz (Department of Pathology, University of Miami School of Medicine, Miami, FL, USA) and graded on glomerular hypercellularity, segmental mesangial expansion, neutrophils/cell debris, crescent formation, and necrosis. These grades were combined for a total glomerular and interstitial pathology score. Deposition of IgG and complement component C3 was analyzed on the half kidney snap frozen in liquid nitrogen. Deposition was assessed by immunofluorescence by incubating slides with rabbit anti-mouse IgG FITC (Sigma) and sheep anti-mouse C3 FITC (Sigma). IgG and C3 were graded 0–3 for intensity of staining, as previously described (4 (link)).
4
Colon Cancer Stem Cell Characterization
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The spherical cells isolated from tumor spheroids of HCT-116 and HT-29 cell cultures without any treatment were firstly verified by their surface markers as we reported before (Wu et al., 2017 (link)). This was to ensure the colon CSCs have been enriched after the sphere culture. For the tumor spheroid culture after each treatment, 1 × 105 spherical cells were used for staining with rabbit anti-human CD133 (prominin-1) antibody (Sigma-Aldrich); mouse anti-human CD44 conjugated with FITC (Invitrogen, Australia); and mouse anti-human CD24 antibody conjugated with RPE (Invitrogen, Australia). For CD133 staining, mouse anti-rabbit IgG-FITC (Sigma-Aldrich) was used as the secondary antibody. After 3 washes with 2% FCS/PBS, the cells were fixed in 2% paraformaldehyde (PFA)/PBS and analyzed by flow-cytometry (Accuri, BD) and CFlow Sampler software. Three biological repeats were performed.
5
Characterization of Cancer Stem Cells
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The spherical cells obtained from sphere culture of HT-29 cells were firstly verified by their surface markers before subsequent experiments. This is to ensure the cancer stem cells have been enriched after sphere culture. The tumour spheres were treated with 1:1 diluted trypsin (2.5% Trypsin-EDTA, Invitrogen, Australia) for 5 min at 37 ° C and washed. Spherical cells were dissociated by repeated pipetting and passing through a cell strainer (40 μM, Falcon, USA). After dissociation, 1 × 105 cells were used for staining with rabbit anti-human CD133 (prominin-1) antibody (Sigma-Aldrich); mouse anti-human CD44 conjugated with FITC (Invitrogen, Australia); and mouse anti-human CD24 antibody conjugated with RPE (Invitrogen, Australia). For CD133 staining, mouse anti-rabbit IgG-FITC (Sigma-Aldrich) was used as the secondary antibody. After 3 washes with 2% FCS/PBS, the cells were fixed in 2% paraformaldehyde/PBS and analysed by flow-cytometry (Accuri, BD) and CFlow Sampler software.
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