Cells were lysed using Triton-X-100 sample buffer, and proteins were separated by SDS-PAGE. Detection occurred by using specific antibodies against β-actin (1:5000 dilution, Biovision through BioCat GmbH, Heidelberg Germany; secondary antibody: IRDye® 800CW Goat anti-Mouse IgG, dilution 1:25,000, Li-Cor Biosciences, Lincoln, NE, USA), SLC35F2 (1:200, Santa Cruz Biotechnology, Dallas, TX, USA; secondary antibody: IRDye® 800CW Goat anti-Mouse IgG, dilution 1:25,000, Li-Cor Biosciences, Lincoln, NE, USA), GAPDH (1:4000, Trevigen via Bio-Techne GmbH, Wiesbaden, Germany; secondary antibody: IRDye® 800CW Goat anti-Rabbit IgG, dilution 1:25,000, Li-Cor Biosciences), ABCB1 (1:1,000, Cell Signaling via New England Biolabs, Frankfurt, Germany; secondary antibody: IRDye® 800CW Goat anti-Rabbit IgG, dilution 1:25,000, Li-Cor Biosciences), p53 (1:1000, Enzo Life Sciences, Lörrach, Germany; secondary antibody: IRDye® 800CW Goat anti-Mouse IgG, dilution 1:25,000, Li-Cor Biosciences), and survivin (1:500, R&D Systems, Minneapolis, MN, USA; secondary antibody: IRDye® 800CW Goat anti-Rabbit IgG, dilution 1:25,000, Li-Cor Biosciences). Protein bands were visualized by laser-induced fluorescence using infrared scanner for protein quantification (Odyssey, Li-Cor Biosciences) and Image Studio Ver. 5.2 software (Li-Cor Biosciences) for densitometric analyses.
Irdye 800cw goat anti mouse igg
Manufactured by LI COR
Sourced in United States, Germany
The IRDye 800CW goat anti-mouse IgG is a fluorescently labeled secondary antibody used for detection in immunological applications. It is designed to bind to mouse primary antibodies, allowing for visualization and quantification of target proteins or molecules.
Automatically generated - may contain errors
Lab products found in correlation
Irdye 680rd goat anti rabbit igg, by LI COR (56 mentions)
Irdye 800cw goat anti rabbit igg, by LI COR (53 mentions)
Odyssey infrared imaging system, by LI COR (43 mentions)
Irdye 680lt goat anti rabbit igg, by LI COR (23 mentions)
Irdye 680 goat anti rabbit igg, by LI COR (16 mentions)
Odyssey imaging system, by LI COR (15 mentions)
Odyssey blocking buffer, by LI COR (12 mentions)
Irdye 680rd goat anti mouse igg, by LI COR (10 mentions)
Odyssey clx imaging system, by LI COR (10 mentions)
Image studio software, by LI COR (10 mentions)
Irdye 680cw goat anti rabbit igg, by LI COR (9 mentions)
Image studio lite, by LI COR (9 mentions)
Odyssey clx, by LI COR (8 mentions)
Nitrocellulose membrane, by Bio-Rad (8 mentions)
Trans blot turbo transfer system, by Bio-Rad (7 mentions)
Odyssey clx infrared imaging system, by LI COR (6 mentions)
β actin, by Merck Group (6 mentions)
Odyssey imager, by LI COR (5 mentions)
Trans blot turbo system, by Bio-Rad (5 mentions)
Anti flag m2 monoclonal antibody, by Merck Group (4 mentions)
220 protocols using irdye 800cw goat anti mouse igg
1
Quantitative Immunoblot Analysis of Cell Signaling Proteins
2
Tricine-SDS-PAGE and Western Blot Analysis
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LPS samples were separated on 14% (w/v) Tricine-SDS-PAGE and the gels stained with silver nitrate (Marolda et al., 2006a) . For Western blot to detect b-O-linked N-acetylglucosamine, samples in the polyacrylamide gel were transferred to nitrocellulose membranes and reacted with O-GlcNAc Monoclonal Antibody (IgM) (Covance) at a 1:1000 dilution. For O-antigen detection anti-O5 antigen monoclonal antibody MF15-4 (Antibodies-online) was used at a 1:10,000 dilution. Alexa Fluor® 680 Goat Anti-Mouse IgM (Invitrogen) or IRDye® 800CW Goat anti-Mouse IgG (LI-COR) were used as secondary antibodies. For protein analysis, polyacrylamide gels were transferred onto nitrocellulose membranes, which were blocked with 10% Western Blocking Solution (Roche Diagnostics). Membranes were incubated overnight at 4 o C with anti-FLAG M2 monoclonal Antibody (Sigma) at a 1:5000 dilution or anti-His (Sigma) at a 1:10,000 dilution. IRDye® 800CW Goat anti-Mouse IgG (LI-COR) or Alexa Fluor® 680 Goat anti-mouse immunoglobulin G (IgG) (Invitrogen) were used as secondary antibodies. Reacting bands were detected by fluorescence with an Odyssey infrared imaging system (Li-cor Bioscience, Lincoln, NE).
3
Western Blot Analysis of Retinol-Binding Protein
4
Western Blot Analysis of Respiratory Syncytial Virus
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Infected Vero cells from single-cycle infections were harvested in 1X NuPage LDS sample buffer (Thermofisher) diluted in PBS containing protease inhibitor (Roche). Cell lysates were homogenized using a QIAshredder spin column (Qiagen) and protein concentrations were determined by BCA assay (Pierce BCA Protein Assay kit). Thirty μg of proteins was denatured in a final composition of 1X NuPage LDS sample buffer (ThermoFisher) and 1X NuPAGE Sample Reducing Agent (Invitrogen) by heating at 90°C for 10 min before being subjected to electrophoresis on NuPAGE 4–15% Mini-PROTEAN TGX Gels (Bio-Rad) with 1X TGS Running Buffer (Bio-Rad). Odyssey Protein Molecular Weight Marker (Li-Cor) was run in parallel. Proteins were transferred to PVDF membranes using the iBlot 2 Gel Transfer Device (ThermoFisher) and stained with a primary mouse anti-RSV F (abcam, ab43812), a mouse anti-RSV P (abcam, ab94965) or a mouse anti-RSV G (abcam, 94966) antibody. The rabbit anti-Tubulin (abcam, ab52866n) antibody was used as a loading control. The secondary antibodies used were goat anti-rabbit IgG IRDye 680RD (at 1:15,000, Li-Cor, 926–68071), and goat anti-mouse IgG IRDye 800CW (1:10,000, Li-Cor, 926–32210). Membranes were scanned using Odyssey software, version 3.0 (Li-Cor).
5
Investigating SKOV-3 Ovarian Cancer Cells
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Human ovarian adenocarcinoma cells (SKOV-3) were purchased from American Type Culture Collection (Manassas, VA, USA). RES was purchased from MilliporeSigma (Burlington, MA, USA). MTT reagent (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was purchased from MilliporeSigma. Guava® Cell Cycle Reagent was purchased from Luminex Corporate (Austin, TX, USA). Annexin V was purchased from ImmunoTools (Friesoythe, Niedersachsen, Germany). Propidium iodide was purchased from MilliporeSigma. Rabbit anti-phospho p38 antibody, rabbit anti p38 antibody, rabbit anti-phospho ERK1/2 antibody, mouse anti-ERK1/2 antibody, rabbit anti-phospho JNK antibody, rabbit anti-JNK antibody, rabbit anti-phospho AKT (Ser473) antibody, mouse anti-AKT antibody, rabbit anti-cyclin A2 antibody, rabbit anti-cyclin B1 antibody, rabbit anti-cyclin E1 antibody, rabbit anti-caspase-9 antibody, rabbit anti-caspase-3 antibody, rabbit anti-PARP antibody, mouse anti-β-actin antibody, and DAPI (4, 6-diamidino-2-phenylindole, dihydrochloride) were purchased from Cell Signaling Technology (Danvers, MA, USA). Goat anti-mouse IgG-IRDye®800CW and goat anti-rabbit IgG-IRDye®680RT were purchased from Li-COR Biosciences (Lincoln, NE, USA). Goat anti-rabbit conjugated with Alexa488 and Goat anti-mouse conjugated with Alexa594 were purchased from Thermo Fisher Scientific (Waltham, MA, USA).
6
Signaling Pathway Characterization
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FACS: Anti-BrdU (1:200, BD Pharmingen). Western Blot: mTOR (1:1000, Cell Signaling), p-mTOR (1:1000, Cell signaling), AKT (1:1000, Cell signaling), p-AKT Ser473 (1:1000, Cell Signaling), ERK1/2 (1:1000, Cell Signaling), p-ERK Thr202/Tyr204 (1:1000, Cell Signaling), AXL (1:10 000, Genscript), p-AXL Tyr702 (1:1000, Cell Signaling) and Tubulin (1:10 000, Sigma).
Secondary antibodies: goat anti-rabbit IgG (IRDye 680RD, 1:10 000) and goat anti-mouse IgG (IRDye 800CW, 1:10 000) were purchased from Li-Cor.
Secondary antibodies: goat anti-rabbit IgG (IRDye 680RD, 1:10 000) and goat anti-mouse IgG (IRDye 800CW, 1:10 000) were purchased from Li-Cor.
7
Quantitative Protein Tag Detection
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A serial dilution of MBP fused to FLAG®-, HA-, myc- and ALFA-tags was prepared in PBS pH7.4, 0.1 µg mL−1 BSA. In total 1 µL of each dilution was spotted on nitrocellulose membranes. Membranes were blocked and washed with 5% milk powder in TBS-T. Established monoclonal antibodies (anti-FLAG® M2, Sigma #F1804; anti-myc 9E10 Synaptic Systems #343 011; anti-HA F-7, SantaCruz #sc-7392) were used in combination with a secondary goat anti-Mouse IgG IRDye800CW (Li-COR #925–32210, dilution 1:500) to detect FLAG®-, myc- and HA-tag, respectively. The ALFA-tag was detected using a FluoTag®-X2 anti-ALFA nanobody (NanoTag Biotechnologies #N1502) directly coupled to IRDye800CW. All primary antibodies and the nanobody were used at 2.7 nM final concentration. Detection of MBP by a rabbit polyclonal serum recognizing MBP (Synaptic Systems, dilution 1:500) and an anti-rabbit IgG IRDye680RD (Li-COR #925–68071, dilution 1:5000) served as an internal loading control. Membranes were scanned using Odyssey CLx (Li-COR). Quantifications were performed using ImageStudioLight (Li-COR).
8
Quantitative Immunoblot Analysis of NF-κB
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Blood was separated into neutrophils and PBMCs. Neutrophils (5 × 106) were used for protein lysates, separated by means of SDS-PAGE, and transferred onto a nitrocellulose membrane. Individual proteins were detected with antibodies against NF-κB p50 (mouse mAb E-10; Santa Cruz Biotechnology, Dallas, Tex), against IκBα (rabbit polyclonal antiserum C-21; Santa Cruz Biotechnology), and against human glyceraldehyde-3-phosphate dehydrogenase (mouse mAb; Merck Millipore, Darmstadt, Germany).
Secondary antibodies were either goat anti-mouse-IgG IRDye 800CW, goat anti-rabbit IgG IRDye 680CW or goat anti-mouse IgG IRDye 680LT (LI-COR Biosciences, Lincoln, Neb). Relative fluorescence quantification of bound secondary antibodies was performed on an Odyssey Infrared Imaging system (LI-COR Biosciences) and normalized to glyceraldehyde-3-phosphate dehydrogenase.
Secondary antibodies were either goat anti-mouse-IgG IRDye 800CW, goat anti-rabbit IgG IRDye 680CW or goat anti-mouse IgG IRDye 680LT (LI-COR Biosciences, Lincoln, Neb). Relative fluorescence quantification of bound secondary antibodies was performed on an Odyssey Infrared Imaging system (LI-COR Biosciences) and normalized to glyceraldehyde-3-phosphate dehydrogenase.
9
Oxyresveratrol Regulates Cell Cycle and Apoptosis in HaCaT Keratinocytes
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Human immortalized keratinocytes (HaCaT) were purchased from Cell Lines Service GmbH (Eppelheim, Germany). Recombinant human TNF-α was purchased from PeproTech (Rocky Hill, NJ, USA). Oxyresveratrol (OXY) was purchased from Sigma-Aldrich (Saint Louis, MO, USA). MTT reagent (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was purchased from Sigma-Aldrich (Saint Louis, MO, USA). Guava® Cell Cycle Reagent was purchased from Luminex Corporate (Austin, TX, USA). Rabbit anti-phospho-AKT (Ser473) antibody, mouse anti-AKT antibody, rabbit anti-phospho-GSK3-β (Ser9), mouse anti-GSK3-β antibody, rabbit anti-Ki-67 antibody, rabbit anti-MCL-1 antibody, DAPI (4′, 6-diamidino-2-phenylindole, dihydrochloride), and LY294002 (a specific inhibitor of the PI3K/AKT) were purchased from Cell Signaling Technology (Boston, MA, USA). Goat anti-mouse IgG-IRDye®800CW and goat anti-rabbit IgG-IRDye®680RT were purchased from Li-COR Biosciences (Lincoln, NE, USA). Goat anti-rabbit conjugated with Alexa488 was purchased from Thermo Fisher Scientific (Waltham (HQ), MA, USA).
10
Western Blot Analysis of Alzheimer's Proteins
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N2a/APP695 cells (1 × 105 cells/mL) were cultured in a 6-well plate for 24 hours and were then treated with RJPs for 24 hours. Cells were washed with cold PBS and lysed with 1 × loading buffer (10 mM Tris, pH 6.8; 1%SDS; 5% glycerin; 0.1 M DTT; bromophenol blue; 1 mM AEBSF (Sigma, USA)). The lysate was collected and boiled at 100 °C for 10 min. Subsequently, an equal volume of sample was electrophoretically separated using 10% SDS-PAGE and then transferred to an NC membrane (Millipore, USA). Membranes were incubated overnight at 4 °C with various primary antibodies. The primary antibodies used in this study: mouseanti-APP (6E10, 1:250, Covance, USA), rabbit-anti-BACE1 (1:500, Abcam, USA), rabbit-anti-HDAC1 (1:5000, AB clonal, Wuhan, China), rabbit-anti-β-actin and rabbit-anti-GAPDH (1:2000, Cell Signaling Technology, USA). The secondary antibodies used were goat-anti-mouse IgG IRDye 800CW (1:10000, Li-COR, USA) and goat-anti-rabbit IgG IRDye 800CW (1:10000, Li-COR, USA). After incubation with the secondary antibody, the membranes were washed 4 times for 5 min each time. Finally, we used the LI-COR Odyssey Infrared Fluorescence Scanning System (Li-COR, USA) to quantify the fluorescence intensity. The intensity of the bands was analyzed using the system software.
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