Anti mouse igg irdye 680lt

Manufactured by LI COR

Sourced in United States

The Anti-mouse IgG IRDye 680LT is a fluorescent labeling reagent designed for use in western blotting, immunohistochemistry, and other protein detection applications. It is an IRDye-labeled secondary antibody that specifically binds to mouse immunoglobulin G (IgG) and can be detected using compatible near-infrared imaging systems.

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Lab products found in correlation

Irdye 800cw anti rabbit igg, by LI COR (4 mentions) Image studio v2, by LI COR (3 mentions) Anti β actin, by Cell Signaling Technology (3 mentions) Gradient polyacrylamide gel, by Bio-Rad (2 mentions) Anti il 2rβ novus, by Novus Biologicals (2 mentions) Anti psmad3 ser423 ser425, by Cell Signaling Technology (2 mentions) Anti tgfβrii, by Cell Signaling Technology (2 mentions) Odyssey infrared imager, by LI COR (2 mentions) Anti smad3, by Cell Signaling Technology (2 mentions) Gradient sds page gel, by Bio-Rad (1 mentions) Odyssey blocking buffer obb, by LI COR (1 mentions) Irdye 800cw goat anti, by LI COR (1 mentions) Halt protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) E coli lps, by Thermo Fisher Scientific (1 mentions) Anti cd14, by Cell Signaling Technology (1 mentions) Anti tlr4, by Santa Cruz Biotechnology (1 mentions) Anti btubulin, by Cell Signaling Technology (1 mentions) Anti p p38 mapk, by Cell Signaling Technology (1 mentions) Anti hur, by Santa Cruz Biotechnology (1 mentions)

5 protocols using anti mouse igg irdye 680lt

Quantitative Western Blotting of Mouse Corneas

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Western blotting was performed as described before.^{22 (link)} Briefly, protein extracts of mouse corneas were prepared in a radioimmunoprecipitation (RIPA) buffer supplemented with 1% SDS and Halt protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific, Waltham, MA, USA). Four corneas were pooled and considered one biological replica. Protein concentration was determined by Bradford-based protein assay (Bio-Rad protein assay). Equal amounts of lysates (30 μg protein) were subjected to electrophoresis in 4% to 15% gradient SDS-PAGE gels (Bio-Rad, Hercules, CA, USA). Protein blots of the gels were blocked with Odyssey blocking buffer (OBB; Li-Cor Biosciences, Lincoln, NE, USA) and incubated overnight with primary antibodies: rabbit anti-α-SMA (ab5694, 1:2000 dilution in OBB; Abcam, Cambridge, MA, USA), goat anti-connective tissue growth factor (CTGF) (clone L-20, 1:500 dilution in OBB; Santa Cruz Biotechnology, Dallas, TX, USA), and mouse anti-β-actin (clone AC-15, 1:10,000 dilution in OBB; Santa Cruz Biotechnology). The secondary antibody used was anti-rabbit IgG IRDye 800CW, anti-goat IRDye 800CW, and anti-mouse IgG IRDye 680LT (1:10,0000 dilution in OBB; Li-Cor Biosciences). Blots were then scanned with the Odyssey Infrared Imaging System, and relative band intensity was quantified by Image Studio v2.0 software (Li-Cor Biosciences).

Chen* W.S., Cao Z., Leffler H., Nilsson U.J, & Panjwani N. (2017). Galectin-3 Inhibition by a Small-Molecule Inhibitor Reduces Both Pathological Corneal Neovascularization and Fibrosis. Investigative Ophthalmology & Visual Science, 58(1), 9-20.

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Western Blot Analysis of Immune Signaling

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Cell lysates were electrophoresed on SDS-PAGE gels (4–15% gradient polyacrylamide gels, BioRad, Hercules, CA, USA) and blots of the gels were probed with the following antibodies: anti-IL-2Rβ Novus, Littleton, CO, USA), anti-TGFβRII, anti-pSTAT5 (Tyr694), anti-STAT5, anti-pSmad3 (Ser423/Ser425), anti-Smad3, and anti-β-actin (all Cell Signaling, Beverly, MA, USA). The secondary Ab used were anti-rabbit IgG IRDye 800CW and anti-mouse IgG IRDye 680LT (LI-COR, Lincoln, NE, USA). Membranes were imaged using the Odyssey Infrared Imager and quantified using Image Studio v2.0 software (LI-COR).

Sampson J.F., Suryawanshi A., Chen W.S., Rabinovich G.A, & Panjwani N. (2015). Galectin-8 promotes regulatory T cell differentiation by modulating IL-2 and TGFβ signaling. Immunology and cell biology, 94(2), 213-219.

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Gal-8 Binding Protein Identification

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To determine whether CD14 or TLR4 are Gal-8 binding proteins, matured BMDMs in 100 mm petri dishes were stimulated with LPS (E. coli-LPS, Invitrogen; 100 ng/mL DMEM, 2 h), washed with PBS, and then lysed in 0.5 mL of RIPA buffer supplemented with a protease inhibitor cocktail. Protein concentration of the lysates were adjusted to 1 mg/mL, 500μg of protein aliquots were precleared by incubation with 50 μl unconjugated agarose beads (2-4 hours at 4°C), supernatants were collected and incubated overnight with Gal-8-conjugated agarose beads (50 μl at 4°C). Then, beads were washed with PBS three times by centrifugation, bound proteins were eluted by boiling the beads in 30μl Laemmli sample buffer for Western blot analysis using anti-TLR4 (1:100 dilution, Santa Cruz,), and anti-CD14 (1:1000 dilution, clone H-4, Cell signaling) as primary antibodies. Secondary antibodies used were anti-rabbit IgG IRDye 800CW and anti-mouse IgG IRDye 680LT (Li-Cor) diluted in OBB (1:10,000, 25°C, 45 minutes). Controls included incubation of lysates with Gal-8 affinity beads in the presence of β-lactose (100 mM, a pan inhibitor of galectins), or sucrose (100 mM, a noncompeting sugar).

Ramadan A., Cao Z., Hassan M., Zetterberg F., Nilsson U.J., Gadjeva M., Rathinam V, & Panjwani N. (2023). Galectin-8 downmodulates TLR4 activation and impairs bacterial clearance in a mouse model of Pseudomonas keratitis. Journal of immunology (Baltimore, Md. : 1950), 210(4), 398-407.

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Western Blot Analysis of Immune Signaling

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Western Blot for Protein Quantification

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Samples were lysed and snap-frozen in RIPA buffer (Sigma) with 1X Halt protease and phosphatase inhibitor cocktail (Invitrogen), loaded onto 4-12% Bis-Tris gels (Novex; Life Technologies) for electrophoresis and then transferred to nitrocellulose membranes for incubation with primary antibodies and secondary antibodies: anti-HuR (Santa Cruz, sc-5261), anti-b-Actin (Cell Signaling Technology, #3700), anti-p-p38 MAPK (Cell Signaling Technology, #4511), anti-b-Tubulin (Cell Signaling Technology, #2146), anti-mouse IgG IRDye 680LT (LICOR Biosciences, #926-68020), anti-rabbit IgG IRDye 800CW (LICOR Biosciences, #926-32211). For ApELAV and p-p38 MAPK experiments, protein levels within each sample was assessed using the LICOR imaging system. Each protein of interest band was normalized to the tubulin or actin within the same sample, and normalized protein of interest in the experimental ganglia was compared to within-animal control ganglia. Data are displayed as fold-expression of normalized protein of interest in experimental sample relative to control sample.

Mirisis A.A., Kopec A.M., & Carew T. (2020). ELAV proteins bind and stabilize C/EBP mRNA in the induction of long-term memory in Aplysia.

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