Log In Sign up
The largest database of trusted experimental protocols

Irdye 800cw anti rabbit igg

Manufactured by LI COR
Visit Supplier
Sourced in United States

The IRDye 800CW anti-rabbit IgG is a near-infrared (NIR) fluorescent dye-labeled secondary antibody. It is designed to detect and quantify rabbit immunoglobulin G (IgG) in various applications.

Automatically generated - may contain errors

Lab products found in correlation

Irdye 680rd anti mouse igg, by LI COR (7 mentions) Gapdh, by Cell Signaling Technology (5 mentions) Odyssey clx, by LI COR (4 mentions) Anti mouse igg irdye 680lt, by LI COR (4 mentions) Anti β actin, by Cell Signaling Technology (4 mentions) Odyssey blocking buffer, by LI COR (3 mentions) Protease inhibitor cocktail, by Merck Group (3 mentions) Odyssey clx infrared imaging system, by LI COR (3 mentions) Irdye 800cw anti mouse igg, by LI COR (3 mentions) Anti β actin, by LI COR (3 mentions) Odyssey infrared imager, by LI COR (3 mentions) Odyssey detection system, by LI COR (3 mentions) Protease and phosphatase inhibitor, by Merck Group (3 mentions) Image studio v2, by LI COR (3 mentions) Odyssey imaging system, by LI COR (2 mentions) Immun blot pvdf membrane, by Bio-Rad (2 mentions) Irdye 680rd anti mouse igg secondary antibodies, by LI COR (2 mentions) Mini protean tris glycine extended gels, by Bio-Rad (2 mentions) Dc protein assay kit, by Bio-Rad (2 mentions) Ripa buffer, by Merck Group (2 mentions)

Spelling variants of this product

IRDye 800CW (808 citations) IRDye 800CW goat anti-rabbit IgG (308 citations) Anti-rabbit IRDye 800CW (80 citations) IRDye 800CW Goat anti-Rabbit IgG Secondary Antibody (52 citations) IRDye 800CW-conjugated goat anti-rabbit IgG (31 citations) IRDye 800CW Donkey anti-Rabbit IgG (H + L) (21 citations) IRDye 800CW goat anti-rabbit IgG (H + L) secondary antibody (4 citations) IgG IRDye 800CW (3 citations) IRDye®800CW anti-rabbit or anti-mouse IgG (2 citations) IRDye 800CW goat anti-rabbit IgG (H+L) antibody (2 citations)

35 protocols using irdye 800cw anti rabbit igg

1

Western Blot Analysis of Muscle Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Muscle homogenates were prepared as previously described (18 (link),25 (link)). Protein was loaded on a 4%–20% gel (Bio-Rad, Mini-PROTEAN TGX Precast Gel) and transferred onto a nitrocellulose membrane overnight at 30 mA at 4°C. Membranes were blocked with 5% non-fat milk for 1 hour and then incubated with primary antibody overnight at 4°C (anti-polyubiquitin, Enzo Life Sciences, Farmingdale, NY, 1:1,000; OXPHOS, MitoSciences, Eugene, OR; α-tubulin, Cell Signaling, Danvers, MA). Membranes were then incubated with secondary antibody for 1 hour (IRDye 800CW anti-Rabbit IgG and IRDye 680RD anti-Mouse IgG; Li-Cor Biosciences, Lincoln, NE). Protein bands were visualized as described above. The amount of poly-ub proteins was normalized to tubulin. Gel-to-gel variation was controlled for by using a standardized sample on each gel.
Standley R.A., Distefano G., Trevino M.B., Chen E., Narain N.R., Greenwood B., Kondakci G., Tolstikov V.V., Kiebish M.A., Yu G., Qi F., Kelly D.P., Vega R.B., Coen P.M, & Goodpaster B.H. (2020). Skeletal Muscle Energetics and Mitochondrial Function Are Impaired Following 10 Days of Bed Rest in Older Adults. The Journals of Gerontology Series A: Biological Sciences and Medical Sciences, 75(9), 1744-1753.
+ Open protocol
+ Expand
2

Western Blotting of Cellular and Exosomal Proteins

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Western blotting was performed as previously described30 (link). Total protein from cells (10 μg) and exosomes (10 μg) was fractionated using an electrophoretic gradient across Mini-PROTEAN tris-glycine extended gels (BIO-RAD, Richmond, CA, USA). Measurement of protein concentration was performed using a Bradford Protein Assay Kit (Takara Bio), according to the manufacturer’s instructions. The gels were then transferred onto Immun-Blot PVDF membranes (BIO-RAD) under wet electrophoretic conditions. The blotted protein was blocked for 1 hr at room temperature with Odyssey blocking buffer in PBS (LI-COR, Lincoln, NE, USA) and was followed by incubation overnight at 4 °C with the following primary antibodies: 1:1000 anti-CD63 mouse monoclonal antibody (BD Biosciences, San Jose, CA, USA); 1:1000 anti-cytochrome-c mouse monoclonal antibody (BD Biosciences); 1:1000 anti-calnexin rabbit monoclonal antibody (Abcam) and 1:1000 hFAB Rhodamine (#12004168 - Bio-Rad). Primary antibodies were detected by IRDye 800CW anti-rabbit IgG and IRDye 680RD anti-mouse IgG secondary antibodies (LI-COR) and were incubated with the protein-blotted membrane for 1 hr at room temperature. Fluorescence was then detected on the Odyssey imaging system (LI-COR).
Morita T., Fujiwara T., Yoshida A., Uotani K., Kiyono M., Yokoo S., Hasei J., Kunisada T, & Ozaki T. (2020). Clinical relevance and functional significance of cell-free microRNA-1260b expression profiles in infiltrative myxofibrosarcoma. Scientific Reports, 10, 9414.
+ Open protocol
+ Expand
3

Purification and Detection of Protein Interactions

Check if the same lab product or an alternative is used in the 5 most similar protocols
The pull-down of endogenously expressed proteins with purified GST-fused proteins was performed using glutathione agarose beads (#745500.10, Macherey-Nagel, Düren, Germany). Beads were coupled to the GST-fused protein for one hour at 4 °C while mixing and centrifuged for 5 min at 500× g. Excess protein was removed by three washing steps. Coupled beads were incubated with HEK293 lysate for one hour at 4 °C on a rotor and again washed 3 times. In the final step, beads were mixed with 1× Laemmli buffer and proteins were denatured at 95 °C for 5 min. Samples were evaluated via SDS-PAGE and western blotting using anti-GST (own antibody, mouse) and anti-IQGAP1 (NBP1-06529, Novus, Wiesbaden Nordenstadt, Germany, rabbit) primary antibodies and secondary antibodies: IRDye® 800 CW anti-Rabbit IgG and IRDye® 680 RD anti-Mouse IgG from LiCor. Values were analyzed by using multiple t test analysis in GraphPad Prism 6 (one unpaired t test per row, fewer assumptions by analyzing each row individually).
Mosaddeghzadeh N., Pudewell S., Bazgir F., Kazemein Jasemi N.S., Krumbach O.H., Gremer L., Willbold D., Dvorsky R, & Ahmadian M.R. (2022). CDC42-IQGAP Interactions Scrutinized: New Insights into the Binding Properties of the GAP-Related Domain. International Journal of Molecular Sciences, 23(16), 8842.
+ Open protocol
+ Expand
4

Western Blot Analysis of TMEM97 and PGRMC1

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in radioimmunoprecipitation assay (RIPA) buffer (Sigma-Aldrich, St. Louis, MO, USA) containing 150 mM NaCl, 1.0% IGEPAL® CA-630, 0.5% sodium deoxycholate, 0.1% SDS, 50 mM Tris, pH 8.0 supplemented with protease inhibitor cocktail, and phosphatase inhibitor cocktail 2 and 3 (Sigma-Aldrich, St. Louis, MO, USA). The cells were sonicated briefly, centrifuged at 13,000×g for 20 min at 4 °C, and the supernatant collected. The protein concentration was determined using a Bio-Rad Dc protein assay kit (Bio-Rad Laboratories, Hercules, CA, USA). Lysates containing 20 µg of protein were run on a 4–20% acrylamide gel and transferred to a PVDF membrane using the Trans-Blot Turbo Transfer System (Bio-Rad Laboratories, Hercules, CA, USA). The PVDF membrane was incubated with Odyssey blocking buffer (Licor Biotechnology, Lincoln, NE) for 1 h at room temperature, then overnight with a rabbit anti-TMEM97 antibody (Aviva Systems Biology, San Diego, CA) at a 1:8000 dilution, or a rabbit anti-PGRMC1 antibody (Sigma-Aldrich) at a 1:1000 dilution at 4 °C, and finally with the secondary antibody, IRDye 800CW anti-rabbit IgG (Licor Biotechnology) at a 1:15,000 dilution. The signals were detected and quantified using the Odyssey® CLx Infrared Imaging System (Licor Biotechnology).
Zeng C., Weng C.C., Schneider ME J.r., Puentes L., Riad A., Xu K., Makvandi M., Jin L., Hawkins W.G, & Mach R.H. (2019). TMEM97 and PGRMC1 do not mediate sigma-2 ligand-induced cell death. Cell Death Discovery, 5, 58.
+ Open protocol
+ Expand
5

HSP60 Knockdown in HEK293 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
HSP60 was knocked down by transfecting 2×106 HEK293 cells with 2 nM of HSP60 Trilencer-27 Human siRNA (OriGene) using Lipofectamine 2000 (Invitrogen). Controls were transfected with siRNA not targeting any human gene (Negative Control siRNA, Origene). Cells were collected 2 d after transfection and knockdown was confirmed by western blot analysis, using 3–8% Tris-Acetate gels (Invitrogen), anti-HSP60 Antibody (Abcam), IRDye® 800CW anti-rabbit IgG (LI-COR), and an Odyssey infrared imaging system [16 (link)]. The expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as control.
Li Y., Malkaram S.A., Zhou J, & Zempleni J. (2014). Lysine biotinylation and methionine oxidation in the heat shock protein HSP60 synergize in the elimination of reactive oxygen species in human cell cultures. The Journal of nutritional biochemistry, 25(4), 475-482.
+ Open protocol
+ Expand
6

Immunofluorescence and Western Blotting Protocols

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The following primary antibodies were used rabbit antiserum R18024 and R18611 [40] (link), rabbit anti-skeletal myosin heavy chain IgG (Santa Cruz Biotechnology), mouse anti-EB1 (BD Biosciences), -β-tubulin (Thermo Scientific), -vinculin (VII-F9, [60] (link)), -GM130 (BD biosciences), -FLAG-M2 antibody (Sigma-Aldrich), and rat anti-FLAG tag (Stratagene). Secondary antibodies used for immunofluorescence studies were highly cross-absorbed Alexa Fluor 488 and 568 anti-rabbit IgG, Alexa Fluor 488 and 568 anti-mouse, and Alexa Fluor 488 anti-rat, all from Invitrogen. For western blotting secondary antibodies were IR-Dye 800cw anti-rabbit IgG (LI-COR) and anti-mouse Alexa 680 (Invitrogen).
Poliakova K., Adebola A., Leung C.L., Favre B., Liem R.K., Schepens I, & Borradori L. (2014). BPAG1a and b Associate with EB1 and EB3 and Modulate Vesicular Transport, Golgi Apparatus Structure, and Cell Migration in C2.7 Myoblasts. PLoS ONE, 9(9), e107535.
+ Open protocol
+ Expand
7

Exosome-induced Signaling Pathway Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were plated in a 96-well plate and treated with 10 μg ml−1 exosomes for 2 h and then processed according to the protocol provided by the manufacturer. In brief, cells were fixed with 4% PFA and washed with 0.1% TritonX-100/PBS. Cells were then blocked using Odyssey blocking buffer for 1 h and stained overnight at 4°C with primary antibody in Odyssey blocking buffer containing 0.1% Tween-20. The next day cells were washed again and incubated with LI-COR secondary antibodies for 1 h at room temperature followed by fluorescent imaging using Odyssey. Antibodies against the following proteins were used: Src (1:100, Cell Signaling; 2109), p-Src (1:100, Cell Signaling; 2101), AKT (1:100, Cell Signaling; 9272), p-AKT (1:100, Cell Signaling; 9271), p38 (1:100, Cell Signaling; 9212), p-p38 (1:100, Cell Signaling; 9211), NF-κB (1:100, Cell Signaling; 3034), p-NF-κB (1:100, Cell Signaling; 3033), NFAT (1:100, Thermo Scientific; PA1-023), ILK (1:100, abcam; ab52480), FAK (1:100, abcam; ab40794) and GAPDH (1:100, Cell Signaling; 14C10). IRDye 800CW anti-rabbit IgG (1:800, LI-COR) were used as secondary antibodies.
Hoshino A., Costa-Silva B., Shen T.L., Rodrigues G., Hashimoto A., Mark M.T., Molina H., Kohsaka S., Di Giannatale A., Ceder S., Singh S., Williams C., Soplop N., Uryu K., Pharmer L., King T., Bojmar L., Davies A.E., Ararso Y., Zhang T., Zhang H., Hernandez J., Weiss J.M., Dumont-Cole V.D., Kramer K., Wexler L.H., Narendran A., Schwartz G.K., Healey J.H., Sandstrom P., Labori K.J., Kure E.H., Grandgenett P.M., Hollingsworth M.A., de Sousa M., Kaur S., Jain M., Mallya K., Batra S.K., Jarnagin W.R., Brady M.S., Fodstad O., Muller V., Pantel K., Minn A.J., Bissell M.J., Garcia B.A., Kang Y., Rajasekhar V.K., Ghajar C.M., Matei I., Peinado H., Bromberg J, & Lyden D. (2015). Tumour exosome integrins determine organotropic metastasis. Nature, 527(7578), 329-335.
+ Open protocol
+ Expand
8

Western Blot Antibody Panel

Check if the same lab product or an alternative is used in the 5 most similar protocols
For western blot (WB), we used the following antibodies: rabbit polyclonal anti‐PAXX/C9orf142 (NovusBio, Littleton, CO, USA; NBP1‐94172, dilution 1 : 1000); rabbit polyclonal anti‐XLF (Bethyl, A300‐730A, 1 : 2000); swine polyclonal anti‐rabbit Ig‐HRP (Dako antibodies, Dako, Glostrup, Denmark; #P0399, 1 : 5000); mouse monoclonal anti‐XRCC4 (NovusBio, NBP1‐48053, 1 : 2000); mouse monoclonal anti‐β‐actin (Abcam, Cambridge, UK; ab8226, 1 : 2000); rabbit polyclonal anti‐mouse Ig‐HRP (Dako antibodies, #P0260, 1 : 5000); goat polyclonal anti‐mouse Ig‐HRP (Dako antibodies, #P0447, 1 : 5000); rat monoclonal anti‐AID (Active Motif, Carlsbad, CA, USA; #39886, 1 : 500) and secondary anti‐rat IgG, HRP‐linked (Cell signaling #7077, Cell Signaling, Leiden, The Netherlands; 1 : 5000); rabbit polyclonal anti‐UNG (custom‐made against murine UNG, 2 μg·mL−1); and IRDye 800CW anti‐rabbit IgG (Licor, Lincoln, NE, USA; #925‐32211, 1 : 15 000).
Dewan A., Xing M., Lundbæk M.B., Gago‐Fuentes R., Beck C., Aas P.A., Liabakk N., Sæterstad S., Chau K.T., Kavli B.M, & Oksenych V. (2018). Robust DNA repair in PAXX‐deficient mammalian cells. FEBS Open Bio, 8(3), 442-448.
+ Open protocol
+ Expand
9

Western Blot Protocol for Protein Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
Samples were diluted in 6xSMASH buffer (375 mM Tris.HCl pH 6.8, 35% Glycerol, 12% SDS, 0.1% bromophenol blue, 4.3 M β-mercaptoethanol), boiled for 10 min at 95 °C, separated on 6% polyacrylamide gel and transferred onto a nitrocellulose membrane by wet transfer in Tris/glycine buffer (100 V for 70 min at 4 °C). Membranes were blocked in TBST buffer (100 mM Tris-HCl pH 7.5, 0.9% NaCl, 0.1% Tween 20, 5% w/v Marvel milk powder) for 1 h at room temperature and then incubated with primary antibodies overnight at 4 °C with gentle rocking. Membranes were washed three times for 10 min with TBST and incubated for 1 h with secondary antibody conjugated to horseradish peroxidase or IRDye 800CW anti-rabbit IgG (Li-COR). After washing three times for 10 min with TBST and one 10 min wash with PBS, bands were visualised either using ECL (GE Healthcare) or on Odyssey Fc Imaging System (Li-COR). Primary and secondary antibody dilutions are listed in Supplementary Table 2.
Gdula M.R., Nesterova T.B., Pintacuda G., Godwin J., Zhan Y., Ozadam H., McClellan M., Moralli D., Krueger F., Green C.M., Reik W., Kriaucionis S., Heard E., Dekker J, & Brockdorff N. (2019). The non-canonical SMC protein SmcHD1 antagonises TAD formation and compartmentalisation on the inactive X chromosome. Nature Communications, 10, 30.
+ Open protocol
+ Expand
10

Antibody-based Autophagy Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies including anti-LC3B (ab48394), anti-SQSTM1 (ab109012), anti-EEA1 (ab2900), anti-Rab5 (ab18211), anti-Rab11a (ab65200), anti-Lamp1 (ab24170), anti-Lamp 2b(ab18529) were purchased from Abcam, GAPDH antibody (60004-1-Ig) was purchased from Proteintech, anti-NSs antibody was generated in our laboratory. E64d (E8640) and pepstatin A (P5318) were purchased from Sigma Aldrich. Conjugated antibodies including the IRDye 800CW goat anti-mouse IgG (926-32210) and IRDye 800CW anti-rabbit IgG (926–32211) were purchased from LI-COR. Fluorescent secondary antibodies including Alexa Fluor 488-conjugated anti-mouse (SA00006-1) and anti-rabbit IgG (SA00006-2), Alexa Fluor 594-conjugated anti-mouse (SA00006-3) and anti-rabbit IgG (SA00006-4) were purchased from Proteintech. DAPI reagent (C0060) was purchased from Solarbio. EGFP-LC3B (11546, deposited by Toren Finkel) was purchased from Addgene. NS-GFP vector was re-constructed by cloning the SFTSV-NSs open reading frame into the pEGFP-N1 vector (preserved in our laboratory). Both siRNA directed against LC3B (GCUUACAGCUCAAUGCUAA) and negative control RNA was synthesized by GenePharma.
Sun Y., Liu M.M., Lei X.Y, & Yu X.J. (2018). SFTS phlebovirus promotes LC3-II accumulation and nonstructural protein of SFTS phlebovirus co-localizes with autophagy proteins. Scientific Reports, 8, 5287.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!