Antibodies used for immunoblots include Rheb (Cell signalling, Cat# 1387, 1:1000), Cre (Cell signalling, Cat# 12830, IHC/IHF - 1:50), Cleaved Caspase 3 (Cell signaling, Cat# 9661, IHC - 1:500), pAkt S473 (Cell signaling, Cat# 4060, 1:1000), pAkt Thr308 (Cell signaling, Cat# 13038, 1:1000), pS6 S240/244 (Cell signaling, Cat# 5364, WB - 1:1000, IHC/IHF – 1:500), Total S6 (Cell signaling, Cat# 2317), p4EBP1 S65 (Cell signaling, Cat# 13443, WB-1:1000, IHC/IHF-1:500), p4EBP1 Thr37/46 (Cell signaling, Cat# 2855, WB-1:1000, IHC/IHF-1:500), 4EBP1 (Cell signaling, Cat# 9644, WB-1:1000), eIF4E (Cell signaling, Cat# 9742, 1:1000), Cyclin D1 (Cell signaling, Cat# 2978, 1:1000), Cyclin D1 (Cell signaling, Cat# 2922, IP-1:50), Cyclin D3 (Cell signaling, Cat# 2936, 1:1000), Actin (Sigma, A5316, 1:5000), PyV mT (clone 762, a gift from Dr. Steven Dilworth, IHC/IHF-1:100), Akt1/2 (Santa Cruz, SC-1619, 1:200), E-Cadherin (BD Bioscience, Cat# 610182, 1:1000), Neu (Dako, A0485, 1:500), Ki67 (Abcam, ab15580, 1:500), Cytokeratin 8/18 (Fitzgerald, 20R-CP004, 1:500), Cytokeratin 14 (BioLegend, PRB-155P, 1:500), and Anti-puromycin (Millipore, Clone 12010 – MABE343, 1:1000). Goat anti-rabbit IRDye 800, Goat anti-mouse IRDye 680, Donkey anti-goat IRDye 800 (Li-COR, 925–32211, 925–68070, 925–68074, 1:10 000)
Goat anti mouse irdye 680
Manufactured by LI COR
Sourced in United States
The Goat anti-mouse IRDye 680 is a secondary antibody conjugated with the IRDye 680 fluorescent dye. It is designed for detection and quantification of mouse primary antibodies in various applications, such as Western blotting, immunohistochemistry, and ELISA.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti rabbit irdye 800, by LI COR (33 mentions)
Odyssey infrared imaging system, by LI COR (13 mentions)
Odyssey blocking buffer, by LI COR (11 mentions)
Irdye 800cw goat anti rabbit, by LI COR (11 mentions)
Goat anti rabbit irdye 680, by LI COR (9 mentions)
Goat anti mouse irdye 800, by LI COR (8 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (6 mentions)
Odyssey imaging system, by LI COR (6 mentions)
Mouse anti tubulin, by Merck Group (5 mentions)
β actin, by Merck Group (4 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (4 mentions)
Odyssey scanner, by LI COR (4 mentions)
Ripa buffer, by Thermo Fisher Scientific (4 mentions)
Odyssey clx imaging system, by LI COR (4 mentions)
Protease inhibitor cocktail, by Thermo Fisher Scientific (4 mentions)
Rabbit anti cas9, by Abcam (3 mentions)
Protease inhibitor cocktail, by Roche (3 mentions)
Odyssey infrared imager, by LI COR (3 mentions)
Odyssey clx, by LI COR (3 mentions)
Thrombin, by Enzyme Research (3 mentions)
75 protocols using goat anti mouse irdye 680
1
Antibody Panel for Immunoblot Analysis
2
Western Blot Analysis of STAT Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
STAT protein expression and phosphorylation status was assessed by western blotting. Cell pellets were lysed in lysis buffer (pH 7.8) containing 50 mM Tris base, 1 mM EDTA, 150 mM NaCl (all Merck), 1% NP40 (Roche Diagnostics), 1 : 100 phosphatase inhibitor cocktail 2 and 3 (Merck), 1X protease inhibitor cocktail, and 1X PhosSTOP (both Roche Diagnostics). Before polyacrylamide gel electrophoresis, Laemmli sample buffer (Bio-Rad) was added 1 : 5 to cell lysates containing 25 μg of protein for STAT1, 3, 5, and 6, 100 μg for pSTAT1, 3, and 6, or 200 μg for pSTAT5. Samples were fractioned by electrophoresis in 8% SDS-PAGE gels, using 30% 37.5 : 1 Acrylamide/Bis solution (Bio-Rad) and further processed for western blot analysis [33 (link)]. After blocking, the primary antibodies in Table 2 were used for overnight staining at 4°C.
After washing, membranes were incubated with 1 : 5000 goat-anti-rabbit IRDye800 (LI-COR Biosciences, order no. 926-32211), 1 : 5000 goat-anti-mouse IRDye680 (LI-COR Biosciences, order no. 926-32220), and 1 : 5000 goat-anti-rat IRDye680 (Invitrogen, order no. A21096) as secondary antibodies and analyzed with the Odyssey Imaging system (LI-COR Biosciences).
After washing, membranes were incubated with 1 : 5000 goat-anti-rabbit IRDye800 (LI-COR Biosciences, order no. 926-32211), 1 : 5000 goat-anti-mouse IRDye680 (LI-COR Biosciences, order no. 926-32220), and 1 : 5000 goat-anti-rat IRDye680 (Invitrogen, order no. A21096) as secondary antibodies and analyzed with the Odyssey Imaging system (LI-COR Biosciences).
3
Quantifying Membrane Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were incubated in vehicle or 1 mM chloroquine (CQ) for 4 hr. Cells were solubilized in RIPA, sonicated, and centrifuged. Supernatants were denatured in sample buffer, run on SDS-PAGE gel, and transferred to polyvinylidene fluoride membrane (PVDF) (Millipore, Bedford, MA). Membranes were immunoblotted for DAT (1:1000) (MAB369; Millipore), β-actin (1:5000) (A5441; Sigma-Aldrich; St. Louis, MO), and Na-K ATPase (1:100; Developmental Studies Hybridoma Bank (DSHB), Iowa City, Iowa). The secondary antibodies used were Li-COR goat anti-rat IRDye 800 (1:15,000), goat anti-rabbit IRDye 680 (1:15,000) and goat anti-mouse IRDye 680 (1:15,000). Band densities were quantified using Image Studio Odyssey Infrared Imaging System (LI-COR, Lincoln, Nebraska).
4
Investigating Phosphatase Regulation of Alpha-Synuclein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Both protease and phosphatase inhibitors were purchased from Roche (Basel, Switzerland). Primary antibodies used in western blotting are as follows: Mouse anti-β-actin (1:5000, Proteintech), Rabbit anti-p-α-syn (1:1000, Abcam), Mouse anti-p-α-syn (1:1000, Wako), Mouse anti-α-syn (1:1000, Santa Cruz), Rabbit anti-Human α-syn (1:2000, Abcam), Mouse anti-demethylate-PP2A (1:1000, Millipore), Mouse anti-PP2A (1:1000, BD biosciences), Mouse anti-β-tubulin (1:1000, Abcam), Mouse anti-PPP2R2A (1:1000, CST), Mouse anti-PPP2R2B (1:1000, Abcam), Rabbit anti-PPP2R2D (1:1000, Abcam), Mouse anti-PPP2R5A (1:500, Santa Cruz), Rabbit anti-PPP2R5D (1:5000, Abcam), Rabbit anti-PPP2R5E (1:1000, Abcam), Mouse anti-LCMT-1 (1:1000, Abcam), Rabbit anti-PME-1 (1:1000, Millipore), Mouse anti-GAPDH (1:3000, Sigma), Mouse anti-c-Myc (1:5000, Clontech), and Mouse anti-FLAG (1:500, Santa Cruz). Secondary antibodies are as follows: Goat anti-Rabbit IRdye680 (LI-COR Bioscience), Goat anti-Mouse IRdye680 (LI-COR Bioscience), Goat anti-Rabbit IRdye800 (LI-COR Bioscience), Goat anti-Mouse IRdye800 (LI-COR Bioscience), Goat anti-Rabbit Alexa Fluor 488 (Thermo Fisher Scientific). The inhibitor of protein phosphatase methylesterase-1 (PME-1), AMZ30, was purchased from MedChem Express (Shanghai, China). Dulbecco’s Modified Eagle’s Medium (DMEM), and fetal bovine serum (FBS) were purchased from Gibco.
5
Western Blot Analysis of Protein Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein extracted using the preceding method was added to reduced Laemmli buffer (Bio-Rad), boiled for 5 min, and loaded into 4%–20% gradient polyacrylamide gels (Bio-Rad). Following electrophoresis, gel contents were transferred to a PVDF membrane (Millipore) and blocked with Odyssey blocking buffer (Li-Cor Biosciences) for 30 min. Membranes were then probed for proteins with the following primary antibodies: SMG1 (Dilution: 1:500; A300-394A, Bethyl Laboratories, Inc.) UPF1 (dilution: 1:1000; anti-RENT1, A301-902A, Bethyl Laboratories, Inc.), Grb2 (dilution: 1:1000; Upstate Cell Signaling), α-Tubulin (1:10,000; Novus Biologicals); and with the following secondary antibodies: goat anti-rabbit IRDye 680 (dilution: 1:10,000, Li-Cor Biosciences), goat anti-mouse IRDye 680 (dilution: 1:10,000, Li-Cor Biosciences), goat anti-mouse IRDye 800 (dilution: 1:10,000, Li-Cor Biosciences). All protein quantification was carried out using Odyssey's Image Studio version 3.0 (Li-Cor Biosciences).
6
Recombinant Glycoprotein B Antigen Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Recombinant glycoprotein B antigen was analyzed by separation of small samples from the purification process on a 10 % SDS-PAGE gel (BioRad Laboratories) and separated products were subsequently transferred to PVDF membrane (Immobilon, Millipore). Gels were run in duplicate; one was Coomassie Blue stained for total protein and the duplicate was analyzed with Western blot using an anti-6His tag mouse monoclonal antibody (Clontech). As a secondary antibody goat anti-mouse IRdye680 (Licor) was used.
Rabbit serum reactivity with recombinant glycoprotein B was detected with Western Blot by incubating the membranes with rabbit serum (1:400 dilution) followed by detection with goat-anti-rabbit IRdye680 (Licor). Similar strategy was done for elephant serum (1:1000 dilution), but primary antibody was detected with rabbit-anti Asian elephant IgG (1:1000 dilution). Sera of rabbits as well as elephants were pre-incubated in buffer containing E.coli lysate (~3 μg/ml) for 1 h at room temperature in order to reduce possible E.coli reactivity. The signal of the conjugate was detected with the Licor Odyssey scanner (Licor).
Rabbit serum reactivity with recombinant glycoprotein B was detected with Western Blot by incubating the membranes with rabbit serum (1:400 dilution) followed by detection with goat-anti-rabbit IRdye680 (Licor). Similar strategy was done for elephant serum (1:1000 dilution), but primary antibody was detected with rabbit-anti Asian elephant IgG (1:1000 dilution). Sera of rabbits as well as elephants were pre-incubated in buffer containing E.coli lysate (~3 μg/ml) for 1 h at room temperature in order to reduce possible E.coli reactivity. The signal of the conjugate was detected with the Licor Odyssey scanner (Licor).
7
Western Blot Analysis of Protein Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were washed with PBS and lysed with ice cold Cell Lysis Buffer (Cell Signaling) supplemented with protease (Roche) and phosphatase inhibitor cocktails (Sigma Aldrich). The bicinchoninic acid method was used for protein quantification using Pierce BCA Protein Assay Kit (Thermo Scientific) according to the manufacturer’s protocol. Lysates were applied to SDS-PAGE (12% SDS-tris glycine) and transferred to 45 µm PVDF membranes (Merck Millipore), which were blocked and stained using Odyssey® Blocking Buffer (Li-Cor). Primary antibodies were incubated overnight at 4°C using a GFP antibody (AnaSpec) and anti-myosin heavy chain (eBioscience) at a dilution of 1:100. Anti-α tubulin (Sigma Aldrich) and anti-Gapdh (Cell Signaling) were used as loading controls (1:5,000). Following washes with TBST buffer, secondary antibodies were incubated for 2 h at RT using goat anti-mouse IRDye680 (Li-Cor Bioscience) and goat anti-rabbit IRDye800 (Li-Cor Bioscience) at 1:10,000 dilutions. Imaging was done with an Odyssey CLx Imaging System (Li-Cor).
8
Antibodies for Extracellular Vesicle Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The primary antibodies used in transmission electron microscopy and Western blotting and ELISA were as follows: mouse anti-CD63 (10628D; Invitrogen, Waltham, MA, USA); mouse-anti CD9 (AHS0902, Invitrogen, Waltham, MA, USA); and mouse anti-ApoB100/48 (3715-3-250, MabTech, Nacka Strand, Sweden). Antibodies anti-CD81 (10630D, Invitrogen, Waltham, MA, USA) and anti-Calnexin (C5C9) (#2679, Cell Signaling Technology, Danvers, MA, USA) were only used on ELISA.
The secondary antibody used in transmission electron microscopy (TEM) was gold-labelled (EM:GAM15, goat anti-mouse 15 nm immunogold-conjugate; BBI Solution, Crumlin, UK). For Western blotting, we used fluorescently labelled secondary antibodies: goat anti mouse IRDye® 800 or goat anti mouse IRDye® 680 or goat anti rabbit IRDye® 680 or goat anti rabbit IRDye® 800 (LI-COR Biosciences, Lincoln, NE, USA). The ELISA secondary antibodies were conjugated with horseradish peroxidase (HRP) as follows: Goat anti rabbit- HRP (111-036-047, Jackson Immunoresearch, West Grove, PA, USA); or Rabbit anti mouse-HRP (JZM035046_Fa, Ancell Immunology Research, Bayport, MN, USA).
The secondary antibody used in transmission electron microscopy (TEM) was gold-labelled (EM:GAM15, goat anti-mouse 15 nm immunogold-conjugate; BBI Solution, Crumlin, UK). For Western blotting, we used fluorescently labelled secondary antibodies: goat anti mouse IRDye® 800 or goat anti mouse IRDye® 680 or goat anti rabbit IRDye® 680 or goat anti rabbit IRDye® 800 (LI-COR Biosciences, Lincoln, NE, USA). The ELISA secondary antibodies were conjugated with horseradish peroxidase (HRP) as follows: Goat anti rabbit- HRP (111-036-047, Jackson Immunoresearch, West Grove, PA, USA); or Rabbit anti mouse-HRP (JZM035046_Fa, Ancell Immunology Research, Bayport, MN, USA).
9
Western Blot Analysis of LRRK1 and LRRK2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh tissues were collected and homogenized in an ice-cold stringent RIPA buffer (50 mM Tris-Cl (pH 7.6), 150 mM NaCl, 0.5 mM EDTA, 1% NP40, 0.5% sodium deoxycholate, 0.1% SDS, 1mM PMSF supplement with protease inhibitor cocktail (P8340, Sigma) and phosphatase inhibitor cocktail (P0044, Sigma)), followed by sonication. Homogenates were centrifuged at 14,000xg for 20 min at 4°C to separate supernatants (RIPA buffer-soluble fraction). An equal amount (10-40 μg per lane) of total proteins from each preparation were loaded and separated on NuPAGE gels (Invitrogen), and transferred to nitrocellulose membranes. The membranes were blocked in Intercept (TBS) Blocking Buffer (927-60001, Li-Cor) for 1 hr at room temperature, and incubated at 4°C overnight with specific primary antibodies. Primary antibodies used were rabbit anti-LRRK1 (ANR-101, Alomone Lab, RRID: AB_2756700), rabbit anti-LRRK2 (ab133474, abcam, RRID: AB_2713963), mouse anti-α-Vinculin (05-386, Millipore, RRID: AB_309711). Membranes were then incubated with dye-coupled secondary antibodies, goat anti-mouse IRdye680 (#925-68070, LI-COR, RRID: AB_2651128) or goat anti-rabbit IRdye800 (#925-32211, LI-COR, RRID: AB_2651127). Signals were quantified using the Odyssey Infrared Imaging System (LI-COR).
10
Quantitative Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cellular proteins were separated using 10% SDS-polyacrylamide gel, electrotransferred onto a polyvinylidene difluoride membrane, and incubated with specific antibodies. The CD300A antibody was purchased from Santa Cruz Biotechnology (CA, USA). Antibodies against GAPDH, AKT, and phosphor-AKT-Ser473 were purchased from Cell Signaling Technology (Shanghai, China). After incubation with the corresponding secondary antibodies, membranes were incubated with goat anti-rabbit IR Dye 800CW or goat anti-mouse IR Dye 680 (LI-COR Biosciences, Lincoln, NE, USA), washed, and scanned for densitometry using an Odyssey infrared imaging system (LI-COR Biosciences, CA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!