Sta 21

Manufactured by Santa Cruz Biotechnology

Sourced in United States

STA-21 is a laboratory instrument designed for the analysis and quantification of proteins and other biomolecules. It utilizes state-of-the-art technology to provide accurate and reliable results. The core function of STA-21 is to perform high-throughput screening and analysis of various biological samples.

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Lab products found in correlation

Fludarabine, by Selleck Chemicals (2 mentions) Ovalbumin (ova), by Merck Group (1 mentions) Mopc 21, by BioXCell (1 mentions) Cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions) Imdm medium, by Merck Group (1 mentions) Digoxin, by Merck Group (1 mentions) Anti mouse ifn γ xmg1, by BioXCell (1 mentions) Anti mouse il 4, by BioXCell (1 mentions) Anti hamster antibody, by MP Biomedicals (1 mentions) Superscript 2, by Thermo Fisher Scientific (1 mentions) Quantstudio 6 flex system, by Thermo Fisher Scientific (1 mentions) Taqman universal pcr master mix, by Thermo Fisher Scientific (1 mentions) Recombinant human il 10, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by PAN Biotech (1 mentions) Facsaria 3, by BD (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Stat3, by Santa Cruz Biotechnology (1 mentions) Dmso control, by Santa Cruz Biotechnology (1 mentions) Lipopolysaccharides (lps), by Santa Cruz Biotechnology (1 mentions) 17β estradiol e2, by Merck Group (1 mentions)

12 protocols using sta 21

Intranasal Asp/Ova Challenge in Mice

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Mice were anesthetized with isoflurane (Piramal, Bethlehem, PA, USA) and were intranasally administered with a mixture of 7 μg of proteinase extracted from Aspergillus melleus (Sigma, St Louis, MO, USA) and 20 μg of Ovalbumin (Ova; Grade V, Sigma, St Louis, MO, USA) (Asp/Ova) in 50 μl of PBS (Katy, TX, USA) every two days for a total of five times (day 0, 2, 4, 6, 8). Sixteen hours after the last challenge, all mice were euthanized and the bronchial lymph nodes, superficial cervical lymph nodes and sera were obtained for further analysis. For TGF-β neutralization experiments, mice were injected intraperitoneally with 200 μg of an anti-TGF-β neutralizing antibody (1D11, BioXCell, West Lebanon, NH, USA) or their corresponding IgG1 control (MOPC-21, BioXCell, West Lebanon, NH, USA) three times every two days (day 0, 2, 4). For STAT3 inhibition experiments, mice were treated with intraperitoneal injections of 0.5 mg/kg STA-21 (Santa Cruz Bio-technology, Santa Cruz, CA, USA) or vehicle every two days for 9 days (day 0, 2, 4, 6, 8) and were treated with intranasal injections of 0.25 mg/kg STA-21 or vehicle every other day for 9 days (day 1, 3, 5, 7).

Choi G, & Chung Y. (2016). Blockade of STAT3 in T Cells Inhibits Germinal Center Reactions against Intranasal Allergens. Biomolecules & Therapeutics, 24(3), 244-251.

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Growth Curves of Cancer Cell Lines

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For growth curve, 5×10⁴ cells/well were seeded in 6-well plates in complete medium or in serum free medium supplemented with 3% WF (SFM-3% WF), in triplicate. Where indicated, S3I-201 (Santa Cruz Biotechnology, Inc.; MDA-MB-231: 50μM; MDA-MB-468: 100μM) or STA-21 (Santa Cruz Biotechnology, Inc., MDA-MB-231 and MDA-MB-468: 30μM) were added to the medium. Fresh medium, with or without inhibitors, was changed every other day. At the indicated times, cells were detached with trypsin-EDTA and counted by Trypan Blue exclusion test.

Segatto I., Berton S., Sonego M., Massarut S., Perin T., Piccoli E., Colombatti A., Vecchione A., Baldassarre G, & Belletti B. (2014). Surgery-induced wound response promotes stem-like and tumor-initiating features of breast cancer cells, via STAT3 signaling. Oncotarget, 5(15), 6267-6279.

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Isolation and Activation of CD4+ T Cells

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The isolation of intestinal lamina proprial cells and flow cytometry were done as previously described(6 (link)). Splenocytes were made into single cell suspension and CD4⁺ T cells were purified with CD4⁺ T cell isolation kit (Miltenyi) with 90–95% purity. 24-well-plates were coated with anti-hamster antibody (MP Biomedical). CD4⁺ T cells were cultured in IMDM medium (Sigma Aldrich) supplied with soluble hamster-anti-mouse CD3 (0.25 µg/ml unless otherwise indicated in the text), hamster-anti-mouse CD28 (1 µg/ml), Gentamicin (50 µg/ml), anti-mouse IL-4 (11B11, 2 µg/ml, BioXCell), and anti-mouse IFN-γ XMG1.2, 2 µg/ml, BioXCell). For iTreg cell differentiation, 5 ng/ml TGF-β was added to the culture. For Th17 cell differentiation, TGF-β was added at 5 ng/ml and IL-6 was added at 20 ng/ml. In some experiments, FICZ was added at a concentration of 200 nM. For blocking RORγt or Stat3 activity, cells were cultured with 10 µM Digoxin (Sigma) or 15 µM STA21 (Santa Cruz), respectively, with controls of DMSO. For IL-21 neutralization, the cells with cultured with 12.5 µg/ml anti-IL21 (R&D Systems) or control IgG.

Heller J.J., Schjerven H., Li S., Lee A., Qiu J., Chen Z.M., Smale S.T, & Zhou L. (2014). Restriction of IL-22-producing T Cell Responses and Differential Regulation of Treg Compartments by Zinc Finger Transcription Factor Ikaros. Journal of immunology (Baltimore, Md. : 1950), 193(8), 3934-3946.

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Regulation of IL-10 and S1P Receptor 1 in NOD Mice

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Five hundred thousand cells from fresh sciatic and lumbar lymph nodes of NOD.Il10^−/− mice were stimulated with recombinant human IL-10 (100 ng/mL from Peprotech), stimulated with IL-10 and STA-21 (10 μM from Santa Cruz Biotech), or incubated without stimulation in XVIVO 15 + Transferrin serum-free media (Lonza) for 30 minutes at 37°C 5% CO₂. RNA was isolated from cells using the Zymo RNA MicroPrep kit. Superscript II (Invitrogen) reverse transcriptase was used to create cDNA. TaqMan universal PCR Master Mix (Applied Biosystems) was used for qPCR. Commercially available TaqMan primer-probe sets for IL-10 and S1Pr1 were used (Applied Biosystems). Cyclophilin A was used as an internal control and detected with the primer-probe set reported by (Su et al., 2008). Reactions were run on a Quantstudio 6 Flex system (Life Technologies) and analyzed as described (Su et al., 2008).

Smith C.J., Allard D.E., Wang Y., Howard JF J.r., Montgomery S.A, & Su M.A. (2018). IL-10 paradoxically promotes autoimmune neuropathy through S1PR1-dependent CD4+ T cell migration.. Journal of immunology (Baltimore, Md. : 1950), 200(5), 1580-1592.

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Enhancing Stem Cell Differentiation

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iSTAT3-MSCs were obtained by adding 10 µM STA21 (Santa Cruz Biotechnology, Dallas, TX, USA) to OA-MSCs for 72 h.

Lee D.H., Kim S.A., Go E.J., Yoon C.Y., Cho M.L., Shetty A.A, & Kim S.J. (2021). Characterization of wild-type and STAT3 signaling-suppressed mesenchymal stem cells obtained from hemovac blood concentrates. Annals of Translational Medicine, 9(16), 1284.

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Isolation and Culture of OA-MSCs

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Adipose tissues were collected from OA patients and were digested with a sterile scissors for 2 min. The tissues were digested with type I collagenase (Cat. LS004197; Worthington Biochemical Products, Lakewood, NJ, USA) at 37°C in a water bath for 40 min and then centrifuged at 1,500 rpm for 5 min. The pellet was resuspended in Dulbecco's phosphate-buffered saline (PBS) and filtered through a 40-μM strainer. Cells were resuspended and cultured with Dulbecco's modified Eagle's medium containing 10% fetal bovine serum (Pan-Biotech, Aidenbach, Germany). The cells were cultured for 4 days until 90% confluent (passage 0) and were then expanded for 2–3 passages and used for experiments. iSTAT3 OA-MSCs were prepared by treatment of OA-MSCs with 10 μM STA21 (Santa Cruz, Texas, USA) for 72 h.

Lee S.Y., Lee S.H., Na H.S., Kwon J.Y., Kim G.Y., Jung K., Cho K.H., Kim S.A., Go E.J., Park M.J., Baek J.A., Choi S.Y., Jhun J., Park S.H., Kim S.J, & Cho M.L. (2018). The Therapeutic Effect of STAT3 Signaling-Suppressed MSC on Pain and Articular Cartilage Damage in a Rat Model of Monosodium Iodoacetate-Induced Osteoarthritis. Frontiers in Immunology, 9, 2881.

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Regulation of Naïve B Cell Fate by STAT and BCL6

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Naïve B cells (CD19⁺IgD^hiCD1d⁻CD27⁻) were isolated from spleens of KPC mice by BD FACS-ARIA III flow cytometry sorting (purity >98%). Sorted naïve B cells were treated with αCD40 (1 μg/mL), LPS (2 μg/mL), rIL-35 (50 ng/mL) with or without BCL6 and STAT inhibitors; STA-21 (20 μmol/L) for STAT3 (Santa Cruz Biotechnology), Fludarabine (50 μmol/L) for STAT1 (Selleckchem) and 79-6 (100 μmol/L) for BCL6 for 72 hrs. The viability and proliferation of naïve B cells and purified Breg cells treated with STAT1 and STAT3 inhibitors were assessed with MTT (Sigma #M5655) as per manufacturer instructions. After 72 hrs, 10 x 10⁶ control or BCL6 and STAT inhibited cells were adoptively transferred via tail vein injection into B cell deficient μMT mice. One day after adoptive transfer, 75,000 KPC4662 cells were orthotopically transplanted into the pancreas of μMT mice. Recipient mice were sacrificed 21 days post-tumor cell injections, tumor size and weight were measured, and spleens and tumors were collected for further processing and analysis.

Mirlekar B., Wang Y., Li S., Zhou M., Entwistle S., De Buysscher T., Morrison A., Herrera G., Harris C., Vincent B.G., Ting J.P., Rashid N., Kim W.Y., Yeh J.J, & Pylayeva-Gupta Y. (2022). Balance between immunoregulatory B cells and plasma cells drives pancreatic tumor immunity. Cell Reports Medicine, 3(9), 100744.

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Induction and Modulation of Collagen-Induced Arthritis

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Induction of CIA was conducted as described previously [13 (link)]. Beginning at the onset of CIA, the mice were dosed orally for 20 days with icariin (25 mg/kg) or an equal volume of water [12 (link)]. In some other experiments, mouse in vivo intraperitoneal injections included recombinant mouse IL-17 rmIL-17 (R&D Systems), 5 μg per mouse for 20 days. Some animals were given 9 consecutive intraperitoneal injections of 0.5 mg/kg STAT3 inhibitor STA-21 (Santa Cruz Biotechnology) in saline, 3 times per week for 3 weeks [14 (link)].

Chi L., Gao W., Shu X, & Lu X. (2014). A Natural Flavonoid Glucoside, Icariin, Regulates Th17 and Alleviates Rheumatoid Arthritis in a Murine Model. Mediators of Inflammation, 2014, 392062.

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Investigating HIV-1 gp120 and Tat Immunomodulation

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PBMCs from HS were treated with HIV-1 gp120, Tat, β-gal (2 μg/ml, ViroGen), or LPS (1 μg/ml, Santa Cruz), in the presence or absence of specific pSTAT-3 inhibitor STA-21 or DMSO control (20 μM, Santa Cruz). The cells were harvested at different time points for staining with CD33-APC, HLA-DR-FITC, CD11b-PE, and CD14-PerCP710, or isotype controls, followed by flow cytometric analysis.

Wang L., Zhao J., Ren J.P., Wu X.Y., Morrison Z.D., Elgazzar M.A., Ning S.B., Moorman J.P, & Yao Z.Q. (2016). Expansion of myeloid-derived suppressor cells promotes differentiation of regulatory T cells in HIV-1+ individuals. AIDS (London, England), 30(10), 1521-1531.

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Inhibitors for Cell Signaling Studies

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17β-Estradiol (E2) and PI3K inhibitor Wortmannin (WM) were purchased from Sigma-Aldrich (Milan, Italy). G-1 (1-[4-(− 6-bromobenzol [1,3]diodo-5-yl)-3a,4,5,9b-tetrahidro3H5cyclopenta[c]quinolin-8yl]-ethanone) and G-15 (3aS, 4R, 9bR)-4-(6-bromo-1, 3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta [c] quinolone were obtained from Tocris Bioscience (Space, Milan, Italy). Src kinase inhibitor PP2 was bought from Selleckchem (DBA, Milan, Italy). MEK inhibitor PD98059 (PD) was purchased from Calbiochem (DBA, Milan, Italy). STAT3 transcription factor signaling inhibitor STA21 and Focal Adhesion Kinase selective inhibitor VS-4718 were bought from Santa Cruz Biotechnology (DBA, Milan, Italy). All the aforementioned compounds were dissolved in dimethyl-sulfoxide (DMSO).

Rigiracciolo D.C., Santolla M.F., Lappano R., Vivacqua A., Cirillo F., Galli G.R., Talia M., Muglia L., Pellegrino M., Nohata N., Di Martino M.T, & Maggiolini M. (2019). Focal adhesion kinase (FAK) activation by estrogens involves GPER in triple-negative breast cancer cells. Journal of Experimental & Clinical Cancer Research : CR, 38, 58.

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