Irdye 680rd donkey anti mouse secondary antibodies

Manufactured by LI COR

Sourced in United States

IRDye® 680RD donkey anti-mouse secondary antibodies are fluorescently labeled antibodies that specifically recognize and bind to mouse primary antibodies. These secondary antibodies can be used in a variety of immunodetection and imaging applications.

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Lab products found in correlation

Ab209780, by Abcam (2 mentions) Irdye 800cw donkey anti rabbit, by LI COR (2 mentions) Pa1 26204, by Thermo Fisher Scientific (2 mentions) Ma5 15122, by Thermo Fisher Scientific (2 mentions) Ma191399, by Thermo Fisher Scientific (2 mentions) Mouse anti β actin, by Proteintech (1 mentions) Odyssey imager, by LI COR (1 mentions) Image studio software, by LI COR (1 mentions) Ab15191, by Abcam (1 mentions) Ab134047, by Abcam (1 mentions) Donkey anti rabbit 800cw, by LI COR (1 mentions) Image studio lite 5, by LI COR (1 mentions) Ab8245, by Abcam (1 mentions)

4 protocols using irdye 680rd donkey anti mouse secondary antibodies

Immunofluorescence analysis of ECM proteins

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Primary antibodies used in this study were: recombinant rabbit anti-cellular communication network factor 2 (CCN2) monoclonal antibody diluted to a concentration of 1:1,000 (Abcam, ab209780); rabbit anti-bone morphogenetic protein 1 (BMP1) polyclonal antibody diluted to a concentration of 1:1,000 (Thermo Fisher Scientific; PA5-103,660; RRID: AB_2852994); rabbit anti-collagen type I (COL1) polyclonal antibody diluted to a concentration of 1:1,000 (Thermo Fisher Scientific; PA1-26204; RRID: AB_2260734); rabbit anti-suppressors of mothers against decapentaplegic homolog 2 (SMAD2) polyclonal antibody diluted to a concentration of 1:1,000 (Thermo Fisher Scientific; 51–1300; RRID: AB_2533896); rabbit anti-phosphorylated SMAD2 (pSMAD2) monoclonal antibody diluted to a concentration of 1:1,000 (Thermo Fisher Scientific; MA5-15122; RRID: AB_10978317); and mouse anti-beta actin (β-actin) monoclonal antibody diluted to a concentration of 1:5,000 (Thermo Fisher Scientific; MA1-91399; RRID: AB_2273656).
For near-infrared fluorescence detection, IRDye® 800CW donkey anti-rabbit and IRDye® 680RD donkey anti-mouse secondary antibodies (LI-COR, 926-32213 and 926-68072, respectively) were used.

Patricelli C., Lehmann P., Oxford J.T, & Pu X. (2023). Doxorubicin-induced modulation of TGF-β signaling cascade in mouse fibroblasts: insights into cardiotoxicity mechanisms. Scientific Reports, 13, 18944.

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Quantification of Extracellular Matrix Proteins

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Primary antibodies used in this study were: recombinant rabbit anti-cellular communication network factor 2 (CCN2) monoclonal antibody diluted to a concentration of 1:1,000 (Abcam, ab209780); rabbit anti-bone morphogenetic protein 1 (BMP1) polyclonal antibody diluted to a concentration of 1:1,000 (Thermo Fisher Scientific; PA5-103660; RRID: AB_2852994); rabbit anti-collagen type I (COL1) polyclonal antibody diluted to a concentration of 1:1,000 (Thermo Fisher Scientific; PA1-26204; RRID: AB_2260734); rabbit anti-suppressors of mothers against decapentaplegic homolog 2 (SMAD2) polyclonal antibody diluted to a concentration of 1:1,000 (Thermo Fisher Scientific; 51-1300; RRID: AB_2533896); rabbit anti-phosphorylated SMAD2 (pSMAD2) monoclonal antibody diluted to a concentration of 1:1,000 (Thermo Fisher Scientific; MA5-15122; RRID: AB_10978317); and mouse anti-beta actin (β-actin) monoclonal antibody diluted to a concentration of 1:5,000 (Thermo Fisher Scientific; MA1-91399; RRID: AB_2273656).
For near-infrared fluorescence detection, IRDye^® 800CW donkey anti-rabbit and IRDye^® 680RD donkey anti-mouse secondary antibodies (LI-COR, 926-32213 and 926-68072, respectively) were used.

Patricelli C., Lehmann P., Oxford J.T, & Pu X. (2023). Doxorubicin-Induced Modulation of TGF-β Signaling Cascade in Mouse Fibroblasts: Insights into Cardiotoxicity Mechanisms. Research Square.

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Quantifying LDH Subunit Expression

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Purified AEC2s were lysed (150 mM NaCl; 50 mM Tris pH 8; 1% NP-40; 1 mM EDTA; 0.5% SDS; 1X protease inhibitor cocktail), then 3 µg of total protein per sample/gel were treated with 50 mM dithiothreitol before SDS-PAGE and western transfer to nitrocellulose as previously described [25 (link)]. Duplicate gels/blots were produced to separately probe with rabbit anti-LDHA or -LDHB antibodies (Proteintech, Rosemont, IL; cat. #s 19987 and 14824). Mouse anti-β Actin (Proteintech, cat #66009) monoclonal was used for both blots. Donkey anti-rab IRDye 800CW and donkey anti-mouse IRDye 680RD secondary antibodies (both LI-COR) were used to simultaneously visualize bands on a LI-COR Odyssey imager, with densitometry and normalization achieved using LI-COR Image Studio software. LDH subunit expression was normalized to β-actin for each sample. Furthermore, to ensure equivalent signals from both LDH antibodies, normalization was also obtained by running purified LDH3 tetramer (Aalto Scientific; composed of 2 LDHA and 2 LDHB subunits) on each gel as a standard.

Newton D.A., Lottes R.G., Ryan R.M., Spyropoulos D.D, & Baatz J.E. (2021). Dysfunctional lactate metabolism in human alveolar type II cells from idiopathic pulmonary fibrosis lung explant tissue. Respiratory Research, 22, 278.

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Protease Hydrolysates Modulate Inflammatory Proteins

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The effect of the treatment with protease hydrolysates on the generation of inflammatory proteins was evaluated after 6 h of TNFα induction EA.hy926 cells. Cells were treated with 2.5 mg/mL of protease hydrolysate for 24 h, and then 10 ng/mL of TNFα was added to the solution for co-treatment for 6 h, and the production of vascular cell adhesion molecule-1 (VCAM-1) and cyclooxygenase 2 (COX2) was determined. The dose of protein hydrolysate was selected based on previous studies [24 (link)]. Cells were lysed using boiling Laemmli buffer containing 50 Mm dithiothreitol (DTT) and 0.2% Triton-X-100. Cell lysates were loaded onto a 9% separating gel and transferred to a nitrocellulose membrane (diameter 0.45 µm, 1620115, Bio-Rad, Montreal, QC, Canada) for incubation with antibodies. Protein bands of VCAM-1 (ab134047, Abcam, ON, Canada), COX2 (ab15191, Abcam) were normalized against glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ab8245, Abcam). Donkey-anti-rabbit 800 CW or donkey-anti-mouse IRDye 680 RD secondary antibodies (Licor Biosciences, Lincoln, NE, USA) were used to visualize the fluorescent protein bands in a Licor Odyssey BioImager, and quantification of protein bands was performed using Image Studio Lite 5.2 (Licor Biosciences, Lincoln, NE, USA).

Li H., Fan H., Lu K., Zhu Q, & Wu J. (2022). Purification of Extracellular Protease from Staphylococcus simulans QB7and Its Ability in Generating Antioxidant and Anti-inflammatory Peptides from Meat Proteins. Nutrients, 15(1), 65.

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