Cells were seeded on coverslips in a 6-well plate, and then exposed to a radiation dose of 6 Gy. After culturing for specified times (0, 0.5, and 24 h), cells were fixed in 4% paraformaldehyde for 20 min, permeabilized with 0.25% Triton X-100 for 40 min, and blocked in 5% bovine serum albumin for 1 h. Cells were immunostained with γ-H2AX (Cell Signaling Technology) at 4 °C overnight, followed by incubation with secondary fluorochrome-labeled antibody (Alexa Fluor 488 donkey anti-rabbit IgG (H + L) antibody; Life Technologies) for 40 min at 37 °C. The cells were then stained with 4′,6-diamidino-2-phenylindole to visualize nuclear DNA. Images were captured using an Olympus fluorescence microscope (Olympus Optical Co., Tokyo, Honshu, Japan).
Alexa fluor 488 donkey anti rabbit igg h l antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) antibody is a secondary antibody conjugated with the Alexa Fluor® 488 fluorescent dye. It is designed to detect and bind to rabbit primary antibodies, specifically targeting the heavy and light chains (H+L) of the immunoglobulin G (IgG) molecule.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Santa Cruz Biotechnology (2 mentions)
Triton x 100, by Merck Group (2 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Fluorescence microscope, by Olympus (1 mentions)
γh2ax, by Cell Signaling Technology (1 mentions)
Anti phospho jak2 tyr1007 1008, by Cell Signaling Technology (1 mentions)
Anti snail, by Santa Cruz Biotechnology (1 mentions)
Anti fibronectin, by Santa Cruz Biotechnology (1 mentions)
Anti vimentin, by Santa Cruz Biotechnology (1 mentions)
Anti mmp 9, by Santa Cruz Biotechnology (1 mentions)
Anti mnsod, by Santa Cruz Biotechnology (1 mentions)
Anti mmp2, by Santa Cruz Biotechnology (1 mentions)
Anti twist, by Santa Cruz Biotechnology (1 mentions)
Anti occludin, by Santa Cruz Biotechnology (1 mentions)
Anti jak1, by Cell Signaling Technology (1 mentions)
Anti jak2, by Cell Signaling Technology (1 mentions)
Anti e cadherin, by Santa Cruz Biotechnology (1 mentions)
Anti n cadherin, by Santa Cruz Biotechnology (1 mentions)
Gsh gssg glo assay kit, by Promega (1 mentions)
13 protocols using alexa fluor 488 donkey anti rabbit igg h l antibody
1
Ionizing Radiation-Induced DNA Damage
2
Leelamine Modulates CXCR7/CXCR4 Signaling
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Leelamine (LEE, Figure 1 A) was purchased from Cayman Chemical (Ann Arbor, MI, USA). LEE stock solution (10 mM) was prepared in EtOH, storage at −20 °C and finally diluted in cell culture medium for use. Anti-CXCR7 and anti-CXCR4 antibodies were purchased from abcam (Cambridge, UK). Anti-MnSOD, anti-fibronectin, anti-vimentin, anti-MMP-9, anti-MMP-2, anti-N-cadherin, anti-E-cadherin, anti-twist, anti-snail, anti-occludin, and anti-b-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-phospho-STAT3(Tyr705), anti-STAT3, anti-phospho-JAK1(Tyr1022/1023), anti-JAK1, anti-phospho-JAK2(Tyr1007/1008), and anti-JAK2 were purchased from Cell Signaling Technology (Beverly, MA, USA). Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) antibody and Alexa Fluor® 594 donkey anti-mouse IgG (H+L) antibody was obtained from Life Technologies (Grand Island, NY, USA). The GSH/GSSG-Glo™ Assay kit was purchased from Promega (Madison, WI, USA).
3
LINC01503 Localization by FISH
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The Cy3-tagged LINC01503 fluorescence in situ hybridization probe was purchased from Ribo Bio (Guangzhou, China). After fixation and permeabilization, cells were incubated with the LINC01503 probe at 42 °C for 12–16 h, and then with anti-SFPQ antibody (Proteintech) at 4 °C overnight. Then, the cells were stained with Alexa Fluor® 488 donkey anti-rabbit IgG (H + L) antibody (Life Technologies), and the nucleus was visualized using Hoechst 33342 (Invitrogen). The digoxin-labeled LINC01503 probe (Supplementary Table 5 ) was purchased from the Boster Biological Technology (Wuhan, China), and the in situ hybridization for tissues was performed according to the manufacturers’ instructions. The images were captured using NIKON ECLIPSE confocal microscope (Niko, Tokyo, Japan).
4
Subcellular Localization of BPIFB1 and Integrins
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5-8F cells transfected or co-transfected with the Flag-BPIFB1 and His-VTN (or -VIM) expression vectors were seeded on coverslips in a six-well plate. After a 24-h culture, cells were fixed in 4% paraformaldehyde for 20 min, subjected to membrane permeabilisation with 0.25% Triton X-100 for 40 min, and blocked in 5% bovine serum albumin for 1 h at room temperature. The cells were then incubated with mouse antibodies against Flag-BPIFB1, ITGAV, VIM and rabbit antibodies against His-VTN/VIM, Src, and N-cadherin antibodies at 4 °C overnight, followed by incubation with secondary fluorochrome-labelled antibodies (Alexa Fluor 488 donkey anti-rabbit IgG (H+L) antibody and Alexa Fluor 568 goat anti-mouse IgG (H+L) antibody; Life Technologies) for 40 min at 37 °C. After incubation with DAPI for 10 min at room temperature to stain the nuclei, cells were imaged using a confocal laser scanning microscope (UltraView Vox; Perkin-Elmer, Waltham, MA, USA). The scatter analysis of channels 561 and 488, as well as the mean relative fluorescence intensity (not total fluorescence intensity), were measured using the image processing and analysis software Velocity (Perkin-Elmer).
5
Immunohistochemistry of Fibroblasts from Skin Biopsies
6
Immunohistochemistry of Fibroblasts from Skin Biopsies
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For immunohistochemistry of fibroblasts from extracted skin biopsies of index case, her parents and control sample were fixed in 3.6% formaldyhyde and washed three times with PBS. Then cells were exposed to 0.3% Triton X-100 (Sigma Chemical) in phosphate buffered saline for 1 minutes and then washed two times with PBS. Cells were blocked in blocking solution containing 1X phosphate buffered saline (PBS), 5% normal bovine serum, 5% normal goat serum, 5% normal donkey serum and 0.1% Tween for half an hour at room temperature. After blocking, specimen were extensively washed in PBS and incubated with primary antibodies diluted in blocking solution on a horizontal shaker overnight at 4°C, and washed in PBS at room temperature for three times. Cells were then incubated with secondary antibodies in blocking solution for 1 hours and washed in PBS for three times at room temperature, then washed and imaged with Zeiss microscopy system. Primary antibody used for immunohistochemistry are: a custom rabbit anti–recombinant mFTO antibody (1:100 dilution). Alexa Fluor®488 Donkey Anti-Rabbit IgG (H+L) Antibody (A-21206, Life technologies) was used as secondary antibody.
7
Ginkgolide C Inhibits Cancer Cell Proliferation
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Ginkgolide C was purchased from Weikeqi Biological Technology (Chengdu, Sichuan, China). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Alexa Fluor® 488 donkey anti-rabbit IgG (H + L) antibody was obtained from Life Technologies (Grand Island, NY, USA). Hepatocyte growth factor (HGF) was purchased from PeproTech (Offenbach, Germany). FITC Annexin V Apoptosis Detection Kit was purchased from BD Pharmingen™ (BD Biosciences, Becton-Dickinson, Franklin Lakes, NJ, USA). iN-fect™ in vitro Transfection Reagent was obtained from iNtRON Biotechnology (Seongnam, Korea). Anti-phospho-c-Met, anti-c-Met, anti-phospho-PI3K(Tyr458), anti-PI3K, anti-phospho-Akt(Ser473), anti-phospho-mTOR(Ser2448), anti-mTOR, anti-phospho-MEK(Ser217/221), anti-MEK, anti-phospho-ERK, anti-ERK, anti-caspase-9, anti-cleaved-caspase-9, and anti-cleaved-caspase-3 antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-Akt, anti-caspase-3, anti-PARP, anti-Bcl-2, anti-Bcl-xL, anti-Survivin, anti-IAP-1, anti-IAP-2, anti-Cyclin D1, anti-COX-2, and anti-β-actin antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
8
Oxymatrine Preparation and Characterization
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Oxymatrine (OMT, Figure 1 A) was purchased from Cayman Chemical (Michigan, USA) and matrine was purchased from Sigma-Aldrich (St. Louis, MO, USA). OMT was stored in a 100 mM stock solution with dimethyl sulfoxide at −20 °C, diluted in the cultured media for in vitro or PBS in vivo experiments. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI), Tris base, glycine, NaCl, sodium dodecylsulfate (SDS), and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). LightShift® Chemiluminescent EMSA kit were purchased from Thermo Fisher Scientific Inc (Waltham, MA, USA). Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) antibody was obtained from Life Technologies (Grand Island, NY, USA). Whole-cell lysates of tumor tissues were obtained with T-PER Tissues Protein Extraction Reagent (Pierce, Rockford, IL, USA).
9
Multicolor Immunocytochemistry of Neuronal Structures
10
Immunostaining of UNC-9 and UNC-44 in C. elegans
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All immunostaining experiments were carried out following the standard protocol using 1-day young adults. The Rabbit anti-UNC-9 antibody was a gift from Dr. Zhao-Wen Wang. The Rabbit anti-UNC-44(L) antibody was a gift from Dr. Anthony Otsuka. Mouse anti-FLAG M2 antibody (Cat# F1804), mouse anti- acetylated tubulin antibody (Cat# T7451) and rabbit anti-GFP (Cat# G1544) were purchased from Sigma. The dilutions for each antibodies are: anti-UNC-9(1:100), anti-UNC-44(1:100); anti-FLAG (1:300), anti-acetylated tubulin(1:300) and anti-GFP (1:150). Alexa Fluor 488 Donkey-anti-rabbit IgG (H+L) antibody (Cat# A-11008) and Alexa Fluor 594 Goat Anti-Mouse IgG (H+L) Antibody (Cat# A-11005) from Molecular Probes were used as secondary antibody in 1:500 dilution.
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