Mouse monoclonal anti stat3

Manufactured by BD

Sourced in United States

The Mouse monoclonal anti-STAT3 is a laboratory reagent used for the detection and quantification of STAT3 protein in various biological samples. STAT3 is a transcription factor that plays a crucial role in cellular signaling pathways. This product can be used in techniques such as Western blotting, immunoprecipitation, and immunohistochemistry to investigate the expression and localization of STAT3 in cells and tissues.

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6 protocols using mouse monoclonal anti stat3

Comprehensive Western Blot Antibody Panel

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In this work we used the following primary antibodies: mouse monoclonal anti-STAT3 (1:1000) (BD Transduction Laboratories, 610189), mouse monoclonal anti-phosphoSTAT3 (1:100) (pY705) (BD Transduction Laboratories, 612356), rabbit polyclonal anti-PARP p85 Fragment pAb (1:500) (Promega, G7341), rabbit polyclonal anti-Mcl-1 (1:500) (Cell Signaling, 5453P) and goat polyclonal anti-caspase 3 (Santa Cruz Biotechnology Inc., sc-1225). To study autophagy we used the following primary antibodies: rabbit polyclonal anti-LC3 (1:1000) (Novus Biologicals, NB100-2220SS), mouse monoclonal anti-p62 (1:1000) (BD Transduction Laboratories, 610832) and rabbit polyclonal anti-Beclin1 (1:1000) (Cell Signaling, 3738S).
Monoclonal mouse anti-β-actin (1:10000) or anti-GAPDH (1:1000) (Santa Cruz Biotechnology Inc., sc-137179) were used as markers of equal loading” in “were used as markers of equal loading. Goat polyclonal anti-mouse IgG-horseradish peroxidase (HRP) (Santa Cruz Biotechnology Inc., sc-2005) rabbit polyclonal anti-goat IgG-HRP (Santa Cruz Biotechnology Inc., sc-2768) and rabbit IgG-HRP (Santa Cruz Biotechnology Inc., sc-2004) were used as secondary antibodies. All the primary and secondary antibodies used in this study were diluted in a PBS- 0.1% Tween 20 solution containing 3% BSA.

Granato M., Gilardini Montani M.S., Filardi M., Faggioni A, & Cirone M. (2015). Capsaicin triggers immunogenic PEL cell death, stimulates DCs and reverts PEL-induced immune suppression. Oncotarget, 6(30), 29543-29554.

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Western Blot Analysis of STAT3, SQSTM1, and LC3

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In western blotting, we used the following primary antibodies: mouse monoclonal anti-STAT3 (1:1000) (BD Transduction Laboratories, San Jose, CA, USA; 612356), mouse monoclonal anti-phosphorylate-STAT3 (Tyr705) (1:100) (Santa Cruz Biotechnology Inc., Heidelberg, Germany; sc-8059), mouse monoclonal anti-sequestome1 (SQSTM1) (1:500) (BD Transduction Laboratories; cat. no. 610833), and rabbit polyclonal anti-LC3 (1:1000) (Novus Biologicals, Cambridge, UK; NB100-2220SS). To assess EBV infection, rabbit polyclonal anti-BRLF1 antibody (1:100) (Bioss Antibodies, Woburn, MA, USA; bs-4542R) and anti-EBNA1 antibody (1:100) (Santa Cruz Biotechnology Inc., Heidelberg, Germany; sc-81581) were used. Mouse monoclonal anti-β-actin (1:10,000) (Sigma Aldrich; A5441) (1:10,000) was used to detect β-actin, serving as a loading control. The goat polyclonal anti-mouse IgG-horseradish peroxidase (HRP, Santa Cruz Biotechnology Inc., Heidelberg, Germany; sc-2005) and anti-rabbit IgG-HRP (Santa Cruz Biotechnology Inc., Heidelberg, Germany; sc-2004) were used as secondary antibodies. All the primary and secondary antibodies were diluted in PBS-0.1% Tween 20 solution containing 3% of BSA (SERVA, Reno, NV, USA; 11943.03).

Granato M., Gilardini Montani M.S., Zompetta C., Santarelli R., Gonnella R., Romeo M.A., D’Orazi G., Faggioni A, & Cirone M. (2019). Quercetin Interrupts the Positive Feedback Loop Between STAT3 and IL-6, Promotes Autophagy, and Reduces ROS, Preventing EBV-Driven B Cell Immortalization. Biomolecules, 9(9), 482.

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Protein Expression Profiling via Immunoblotting

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To evaluate the expression of proteins we used the following antibodies: mouse monoclonal anti-STAT3 (1:500) (BD Transduction Laboratories, 610,189), rabbit polyclonal anti-phospho-STAT3 (1:500) (p-Tyr705, clone D3A7, Cell Signaling Technology, 9145), mouse monoclonal anti-p53 (1:100) (clone DO-1, Santa Cruz Biotechnology Inc., sc-126), mouse monoclonal anti-HSP90 (1:100) (Santa Cruz Biotechnology Inc., sc-69703), mouse monoclonal anti-p21 (1:100) (clone F-8, Santa Cruz Biotechnology Inc., sc-271610), mouse monoclonal anti-SREBP1 (1:100) (clone A-4, Santa Cruz Biotechnology Inc., sc-365513), mouse monoclonal anti-MVK (1:100) (clone D-3, Santa Cruz Biotechnology Inc., sc-390669). Mouse monoclonal anti-β-actin (1:10,000) (Novus Biological, NB600-501) and goat polyclonal anti-lamin B (1:100) (Santa Cruz Biotechnology Inc, sc-374015) were used as loading control. The goat anti-mouse IgG-Horseradish Peroxidase (Santa Cruz Biotechnology Inc., sc- 2005), goat anti-rabbit IgG-HRP (Santa Cruz Biotechnology Inc., sc-2004) and rabbit anti-goat IgG-HRP (Santa Cruz Biotechnology Inc., sc-2768) were used as secondary antibodies. All the primary and secondary antibodies were diluted in PBS-0.1% Tween20 solution containing 3% of BSA (SERVA).

Romeo M.A., Gilardini Montani M.S., Benedetti R., Santarelli R., D'Orazi G, & Cirone M. (2020). STAT3 and mutp53 Engage a Positive Feedback Loop Involving HSP90 and the Mevalonate Pathway. Frontiers in Oncology, 10, 1102.

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Western Blot Analysis of KSHV Infection

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In all, 1 × 10⁶ uninfected or KSHV-infected cells were lysed, subjected to electrophoresis and transferred to nitrocellulose membranes, as previously described.^{29 (link)} Membranes were blocked in PBS-0.1% Tween 20 solution containing 3% BSA, probed with specific antibodies and developed using ECL Blotting Substrate (Advansta). The following antibodies were used: mouse monoclonal antibody against Kb-ZIP (Santa Cruz Biotechnology, sc-69797), p STAT6 (1:100; Santa Cruz Biotechnology Inc., sc-136019), STAT6 (1:100; Santa Cruz Biotechnology Inc., sc-1689), mouse monoclonal anti-STAT3 (1:1000; BD Transduction Laboratories, 610189), mouse monoclonal anti-phospho-STAT3 (p-Tyr705, 1:100; Santa Cruz Biotechnology Inc., sc-8059), p STAT1 (1:100; Santa Cruz Biotechnology Inc., sc-136229), STAT1 (1:100; Santa Cruz Biotechnology Inc., sc-464), mouse monoclonal anti-Ire1α (1:100; Santa Cruz Biotechnology, sc-390960), XBP1s (NovusBio NBP1-77681SS), ATF4 (R&D system, MAB7218), rabbit polyclonal anti-BIP (1:100; Cell Signaling, 3177), mouse monoclonal anti-CHOP (1:100; Santa Cruz Biotechnology, sc-7351), and anti-β-actin (1:10000; Sigma Aldrich, A5441). Goat anti-mouse IgG-HRP and anti-rabbit IgG-HRP (1:10.000 Santa Cruz Biotechnology Inc) were used as secondary antibodies.

Gilardini Montani M.S., Falcinelli L., Santarelli R., Granato M., Romeo M.A., Cecere N., Gonnella R., D’Orazi G., Faggioni A, & Cirone M. (2020). KSHV infection skews macrophage polarisation towards M2-like/TAM and activates Ire1 α-XBP1 axis up-regulating pro-tumorigenic cytokine release and PD-L1 expression. British Journal of Cancer, 123(2), 298-306.

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Comprehensive Immunoblot Profiling

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The following primary antibodies were used: rabbit polyclonal anti-IL-6 (Gene Tex, Alton Pkwy Irvine, CA), rabbit polyclonal anti-IL-1β (Gene Tex, CA, USA), mouse monoclonal anti-TNFα (Santa Cruz), rabbit polyclonal anti-HMGB1 (Abcam, Cambridge, UK), mouse monoclonal anti-granzyme B (Calbiochem, San Diego, CA, USA), mouse monoclonal anti-phospho-STAT3 (pY705) (BD Transduction Laboratories, San Jose, CA, USA), mouse monoclonal anti-STAT3 (BD Transduction Laboratories), rabbit polyclonal anti-LC3 (Novus Biologicals, Littleton, CO, USA), mouse monoclonal anti-p62 (BD Transduction Laboratories) and mouse monoclonal anti-β-actin Ac-40 (Sigma-Aldrich).

D'Eliseo D., Pannucci E., Bernini R., Campo M., Romani A., Santi L, & Velotti F. (2019). In vitro studies on anti-inflammatory activities of kiwifruit peel extract in human THP-1 monocytes. Journal of ethnopharmacology, 233.

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Immune Signaling Protein Detection

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The following primary antibodies were used: rabbit polyclonal anti-IL-6 (Gene Tex, Alton Pkwy Irvine, CA), rabbit polyclonal anti-IL-1β (Gene Tex, CA, USA), mouse monoclonal anti-TNFα (Santa Cruz), rabbit polyclonal anti-HMGB1 (Abcam, Cambridge, UK), mouse monoclonal antigranzyme B (Calbiochem, San Diego, CA, USA), mouse monoclonal anti-phospho-STAT3 (pY705) (BD Transduction Laboratories, San Jose, CA, USA), mouse monoclonal anti-STAT3 (BD Transduction Laboratories), rabbit polyclonal anti-LC3 (Novus Biologicals, Littleton, CO, USA), mouse monoclonal anti-p62 (BD Transduction Laboratories) and mouse monoclonal anti--actin Ac-40 (Sigma-Aldrich).

Wang C., Yan L., Yan B., Zhou L., Sun W., Yu L., Liu F., Du W., Yu G., Hu Z., Yuan Q., Xiao L., Li H., Tong P., Zhang J., Shan L, & Efferth T. (2019). Agkistrodon ameliorates pain response and prevents cartilage degradation in monosodium iodoacetate-induced osteoarthritic rats by inhibiting chondrocyte hypertrophy and apoptosis. Journal of ethnopharmacology, 231.

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