For immunofluorescence MitoTracker-stained cells were fixated, residual PFA was quenched with NH4Cl (50 mM) in PBS for 30 min, cells were permeabilized with Triton X-100 (0.5% v/v) and, after washing, blocked according to the manufacturer’s instructions. The cells were covered with the antibody (Drp1: Cell Signalling, #8570, 1:50 dilution, 1 h at RT; α-Tubulin, Sigma Aldrich, St. Louis, Missouri, USA, T5168, 1:500, 1 h at RT), washed and incubated with the secondary antibody (Anti-Rabbit IgG-FITC, Sigma-Aldrich, #F9887, 1:100, 45 min at RT; anti-Mouse IgG-FITC, Sigma-Aldrich, F9137, 1:100, 1 h at RT), then mounted and analysed by fluorescence microscopy (λexc. = 470 ± 20 nm, λem. = 525 ± 25 nm).
Anti rabbit igg fitc
Manufactured by Merck Group
Sourced in United States
Anti-rabbit IgG-FITC is a fluorescently labeled secondary antibody that binds to rabbit immunoglobulin G (IgG). It is commonly used in immunoassays and immunofluorescence techniques to detect and visualize rabbit primary antibodies.
Automatically generated - may contain errors
Lab products found in correlation
Anti mouse igg fitc, by Merck Group (6 mentions)
Anti sars cov 2 polyclonal s1 antibody, by Sino Biological (4 mentions)
Attune nxt flow cytometer, by FlowJo (3 mentions)
Anti mouse igg tritc, by Merck Group (3 mentions)
Optiphot diginet camera system, by Nikon (2 mentions)
Immobolin crescendo western hrp substrate, by Merck Group (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
α tubulin, by Merck Group (1 mentions)
Cryostat microtome, by Leica (1 mentions)
In situ cell death detection kit with fluorescein, by Roche (1 mentions)
Amersham imager 600, by GE Healthcare (1 mentions)
Anti mouse igg fitc, by Abcam (1 mentions)
Rabbit anti human tyrosine hydroxylase th, by Abcam (1 mentions)
Anti rabbit igg tritc, by Abcam (1 mentions)
Rabbit anti human lmx1a, by Abcam (1 mentions)
Rabbit anti human nurr1, by Santa Cruz Biotechnology (1 mentions)
Hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Anti ki67, by Santa Cruz Biotechnology (1 mentions)
Anti glucagon, by Santa Cruz Biotechnology (1 mentions)
Anti insulin, by Santa Cruz Biotechnology (1 mentions)
26 protocols using anti rabbit igg fitc
1
Immunofluorescence Imaging of Mitochondrial Dynamics
2
Quantifying SARS-CoV-2 S Protein Expression
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HEK293T cells used to produce lentiviral vectors were harvested 72-hours post-transfection. Cells were incubated in PBS+5mM EDTA for 7 minutes at 37°C to mediate disassociation. The cells were fixed in 3.7% formaldehyde and stained with anti-SARS-CoV-2 polyclonal S1 antibody (Sino Biological, 40591-T62; RRID: AB_2893171) and secondary antibody anti-Rabbit-IgG-FITC (Sigma, F9887, RRID: AB_259816). S surface expression was measured using a Life Technologies Attune NxT flow cytometer and data was processed using FlowJo v7.6.5 (Ashland, OR).
3
Immunohistochemical Analysis of Rat Brain
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Staining of slices was performed as previously described (Wang et al., 2019 (link)). Briefly, rats were deeply anesthetized and perfused transcardially with ice-cold 4% paraformaldehyde. Rat's brain was post-fixed with 4% paraformaldehyde for 12 h, dehydrated with 30% sucrose/phosphate-buffered saline (PBS) at 4°C for 3 days, and then cut into 10-μm thickness coronal slices (−2.5 to −5 mm from bregma) using a Leica cryostat microtome. Coronal slices were permeabilized with 0.5% Triton X-100 and blocked with 5% goat serum, incubated with anti-NeuN rabbit antibody (1:200, #12943, CST), anti-NeuN mouse antibody (1:200, #94403, CST), anti-HSP27 (1:200, ab2790, Abcam), anti-Iba-1 (1:200, #19741, Wako) primary antibody at 4°C overnight and then incubated with the Anti-Mouse IgG-TRITC (1:100, T5393, Sigma) or Anti-Rabbit IgG-FITC (1:100, F9887, Sigma) at room temperature for 2 h. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining was assayed using in situ Cell Death Detection Kit with Fluorescein (11684795910, Roche) according to the manufacturer's instruction. Images were captured using a fluorescence microscope (BX51, Olympus) and analyzed with Image J software.
4
Immunocytochemistry Protocols for Enpp4
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Immunocytochemistry was carried out on fixed unpermeabilized cells with polyclonal antibodies anti-XlEnpp4 used at 1/200 or anti-mEnpp4 (CR65; see ref. 31 (link)) used at 1/400 and anti-rabbit IgG FITC (Sigma) at 1/80. The staining was recorded using a Nikon Optiphot/ Diginet camera system. Photographs were taken at a magnification of ×40.
5
SARS-CoV-2 Spike Protein Western Blot
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The remaining virus producer cells leftover after taking cells for flow cyometry were lysed using RIPA buffer supplemented with protease inhibitor cocktails (RIPA: 50 mM Tris pH 7.5, 150 mM NaCl, 1 mM EDTA, Nonidet P-40, 0.1% SDS, PI + PMSF: Sigma, P8340) for 40 min on ice. Lysate was then harvested and used for western blotting. Samples were separated using a 10% acrylamide SDS-PAGE gel and transferred to a PVDF membrane. Blots were then incubated with polyclonal anti-S2 antibody (Sino Biological, 40590; RRID:AB_2857932), anti-S1 antibody (Sino Biological, 40591-T62; RRID: AB_2893171), and anti-GAPDH as a loading control (Santa Cruz, Cat# sc-47724, RRID: AB_627678). Secondary antibodies used were anti-Rabbit-IgG-FITC (Sigma, A9169; RRID: AB_258434) and anti-Mouse-IgG-FITC (Sigma, Cat# A5278, RRID: AB_258232). Blots were imaged using Immobolin Crescendo Western HRP substrate (Millipore, WBLUR0500) and exposed on a GE Amersham Imager 600. Quantification of band intensity was determined using ImageJ (NIH, Bethesda, MD).
6
Immunofluorescence Characterization of hADSCs
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hADSCs were differentiated in 24-well chambers at 29 days, rinsed three times with PBS, and incubated in 4% paraformaldehyde overnight at 4°C. Bovine serum albumin (1%) in PBS was used for blocking. Afterwards, the cells were incubated overnight at 4°C with the following primary antibodies: mouse anti-human Cav-1 (1:100; Cell Signaling Technology), rabbit anti-human tyrosine hydroxylase (TH) (1:100; Abcam), rabbit anti-human Lmx1a (1:100; Abcam) or rabbit anti-human Nurr1 (1:100; Santa Cruz Biotechnology). The samples were then rinsed three times thoroughly with PBS and incubated with the following secondary antibodies: anti-rabbit IgG-FITC (Sigma-Aldrich), anti-mouse IgG-FITC (Abcam), anti-mouse IgG-TRITC (Sigma-Aldrich) or anti-rabbit IgG-TRITC (Abcam) for 1.5 hours at room temperature. The cells were thereafter rinsed three times in PBS and incubated for 5 minutes with Hoechst 33258. The samples were then washed twice with PBS and once in deionized water. Stained cells were observed under a confocal laser scanning microscope (SP8, Leica).
7
Quantitative Analysis of Pancreatic Islet Morphology
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Pancreatic and liver tissues were fixed with 10% formaldehyde and paraffin embedded by standard methods. Subsequently, the tissues were sectioned (5 µm thin) and stained with Hematoxylin & Eosin (H & E) and were mounted by mounting medium. At least 5 islets in each section were randomly chosen and analyzed. The cell number of each islet was quantified with Image J software (https://imagej.nih.gov/ij/ ). Tissue morphology was examined and analysed by clinical pathologist.
Pancreatic tissue sections were subjected to immunohistochemistry for insulin and glucagon using anti-Insulin (Santa Cruz Biotechnology USA; sc- 25840), anti-glucagon (Santa Cruz biotechnology USA; sc-74825) and anti-Ki-67 (Santa Cruz biotechnology USA; sc-101861) antibodies and HRP conjugated secondary antibody (Santa cruz Biotechnology USA; sc-2030) as per standard protocols. The sections were counter stained with Harris Hematoxylin (Nice Co., India) and the detection was based on the intensity of brown chromogen produced by HRP-DAB at the site of enzyme activity (3, 3′-diaminobenzidine) and the images were captured using light microscope. Subsequently for fluroscence imaging, anti-rabbit IgG-FITC (Sigma-Aldrich Inc., USA; F6005) and anti-mouse IgG-TRITC (Sigma-Aldrich Inc., USA; T5393) were used.
Pancreatic tissue sections were subjected to immunohistochemistry for insulin and glucagon using anti-Insulin (Santa Cruz Biotechnology USA; sc- 25840), anti-glucagon (Santa Cruz biotechnology USA; sc-74825) and anti-Ki-67 (Santa Cruz biotechnology USA; sc-101861) antibodies and HRP conjugated secondary antibody (Santa cruz Biotechnology USA; sc-2030) as per standard protocols. The sections were counter stained with Harris Hematoxylin (Nice Co., India) and the detection was based on the intensity of brown chromogen produced by HRP-DAB at the site of enzyme activity (3, 3′-diaminobenzidine) and the images were captured using light microscope. Subsequently for fluroscence imaging, anti-rabbit IgG-FITC (Sigma-Aldrich Inc., USA; F6005) and anti-mouse IgG-TRITC (Sigma-Aldrich Inc., USA; T5393) were used.
8
Comprehensive Antibody Validation Protocol
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The following antibodies were used in this study: anti‐HA (ab9110), anti‐FLAG (ab1162), anti‐SC‐35 (ab11826), anti‐H3K9me3 (ab8898), anti‐METTL3 (ab66660), and anti‐p16 (ab54210) were purchased from Abcam; anti‐METTL14 (PA5‐58204) was from Thermo Fisher Scientific; anti‐mouse IgG‐Cy3, anti‐rabbit IgG‐Cy3, anti‐mouse IgG‐FITC, anti‐rabbit IgG‐FITC, anti‐FLAG M2 affinity gel, and monoclonal anti‐HA agarose were from Sigma‐Aldrich; anti‐Lamin A/C (sc‐20681), anti‐Lamin A (sc‐518013), anti‐Lamin A (sc‐71481), anti‐p21 (sc‐6246), and anti‐p16 (ab54210) were from Santa Cruz Biotechnology; anti‐m6A (No. 202003) was from Synaptic Systems; rabbit anti‐γH2AX (05‐636) was from EMD Millipore; rabbit anti‐HMGB1 (#3935), anti‐Ubiquitin (#3936), and GAPDH (14C10) mAb (#2118) were from Cell Signaling Technology; and mouse/rabbit Alexa Fluor 488/Alexa Fluor 594 was from Thermo Fisher Scientific.
9
Bacterial Strains and Reagents for Molecular Biology
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All chemical reagents were molecular biology grade. Escherichia coli TG1 was obtained from MRC, Cambridge, and used for cloning and amplification of phages. E. coli HSM174 pLysS (Novagen) was used for protein expression. The anti-M13/HRP and His prob-HRP detection kits were purchased from Amersham-Pharmacia Biotech (Uppsala). Protein L peroxidase-HRP, anti-rabbit IgG-FITC and Calcofluor white stain were purchased from Sigma. Bradyrhizobium strain DOA9 and other bradyrhizobia were isolated from A. americana in our laboratory [1 (link)], while the reference strains of B. diazoefficiens USDA110 and Bacillus subtilis 168 were obtained from the School of Biotechnology, SUT.
10
Cytological Analysis of Plant Meiosis
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Fixation of flower buds, slide preparation of PMCs, and fluorescence in situ hybridization (FISH) were carried out as previously described27 (link). Three plants of each mutant and their corresponding wild types were analyzed. Squash preparations were made removing gynoecia from fixed flowers. The gynoecia were transferred to a slide, stained with acetic carmine and squashed. Immunolocalization of modified histone H3 was carried out as previously described, with some modifications22 (link). Immunolocalizations of ASY1 (1:1000), ZYP1 (1:500) and DMC1 (1:250) proteins were carried out as previously described68 (link) with the primary antibodies shown in Supplementary Table S5 . The secondary antibodies were anti-rabbit IgG FITC conjugated (1:50, Sigma) and anti-rat IgG Cy3 conjugated (1:50, Sigma).
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