Anti akt1 2 3

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-AKT1/2/3 is a laboratory reagent that specifically recognizes and binds to the AKT1, AKT2, and AKT3 proteins. This product is commonly used in research applications to detect and analyze the expression levels of these proteins.

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Lab products found in correlation

Anti p akt1 2 3, by Santa Cruz Biotechnology (3 mentions) Anti β actin, by Merck Group (2 mentions) Enhanced chemiluminescence kit, by Santa Cruz Biotechnology (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Anti trka, by Santa Cruz Biotechnology (2 mentions) Anti ptrka, by Santa Cruz Biotechnology (2 mentions) Dulbecco s modified eagle s medium, by Cytiva (2 mentions) Poly d lysine, by BD (2 mentions) Anti β actin antibody, by Abcam (2 mentions) Dc protein assay, by Bio-Rad (2 mentions) Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions) Anti e cadherin, by BD (1 mentions) Anti ki67, by Merck Group (1 mentions) Anti n cadherin, by BD (1 mentions) Scintillation cocktail, by Thermo Fisher Scientific (1 mentions) Tetrachloroauric acid trihydrate, by Merck Group (1 mentions) Anti vimentin, by BD (1 mentions) Anti β catenin, by BD (1 mentions) Anti α sma, by Merck Group (1 mentions) Anti cd31, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

Akt1/2/3 (24 citations) Anti-Akt1 (15 citations) Anti-p-Akt1/2/3 (10 citations) Akt1/2 (8 citations) Rabbit anti-Akt1/2/3 (3 citations) Anti-AKT1/2/3 (H-136, (3 citations) Anti–p-Akt1/2/3 (sc-7985-R), (2 citations) Rabbit anti-p-AKT1/2/3 (Ser473) polyclonal antibody (sc-101629) (2 citations)

9 protocols using anti akt1 2 3

Evaluation of Cancer Cell Behavior

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Tetrachloroauric acid trihydrate, trisodium citrate and sodium borohydride were from Sigma-Aldrich, St. Louis, MO. [³H] Thymidine was from Perkin-Elmer, (Waltham, MA). Media and PBS was purchased from Mediatech (Manassas, VA). Cisplatin was obtained from the Mayo Clinic Pharmacy services at a concentration of 50mg/ml. Scintillation cocktail was purchased through Fisher Scientific. And Alexa Fluor® 488 Phalloidin is from Life Technologies.
The following antibodies were used for Western blotting and immunofluorescence: anti–E-cadherin, anti-N-Cadherin, anti-β-Catenin, and anti-vimentin (BD Biosciences); anti-α-SMA, anti-Ki67, and anti-β-actin (Sigma-Aldrich); anti-IκBα and anti-p65 (Cell Signaling Technology); anti-CD31, anti-AKT1/2/3, and anti-phos-AKT1/2/3 (Santa Cruz Biotechnology); anti-NUP214 (Bethyl Laboratories, Inc.) Secondary antibodies were from Santa Cruz Biotechnology, Inc.

Xiong X., Arvizo R.R., Saha S., Robertson D.J., McMeekin S., Bhattacharya R, & Mukherjee P. (2014). Sensitization of ovarian cancer cells to cisplatin by gold nanoparticles. Oncotarget, 5(15), 6453-6465.

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Western Blot Analysis of Macrophage Proteins

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Macrophages were homogenized in cold RIPA lysis buffer containing protease inhibitor cocktails (Sigma; 4693116001) and phosphatase PhosStop EASYPack cocktails (Roche; 4906837001) followed by sonication with a tip-probe sonicator on ice. The concentrations of the proteins were determined using the bicinchoninic acid (BCA) assay. Equal amounts of proteins were loaded onto 10% or 12% SDS‒PAGE gels, separated by electrophoresis, and then transferred onto polyvinylidene fluoride (PVDF) membranes. The membranes were subsequently incubated with primary antibodies. The protein bands were visualized using an enhanced chemiluminescence (ECL) system (Amersham Biosciences, Cytiva) and imaged using an iBright FL1500 Imaging System (Thermo Fisher). The primary antibodies used were anti-ARG1 (1:1000, sc-47715), p-ribosomal protein S6 (1:1000, sc-293144), anti-ribosomal protein S6 (1:1000, sc-74459), and anti-p-4E-BP1 (1:1000, sc-293124); anti-4E-BP1 (1:1000, sc-81149); anti-p-AKT1/2/3 (1:1000, sc-81433); and anti-AKT1/2/3 (1:1000, sc-81434) obtained from Santa Cruz; and anti-PHGDH (1:750, 13428), anti-CCR2 (1:1000, 12199) obtained from CST; and anti-α-Tubulin (1:1000, 1224-1-AP) and secondary antibodies horseradish peroxidase (HRP)-conjugated anti-rabbit Ig (H + L) (1:5000, SA00001-2) and HRP-conjugated anti-mouse Ig(H + L) (1:5000, SA00001-1) were purchased from Proteintech.

Cai Z., Li W., Hager S., Wilson J.L., Afjehi-Sadat L., Heiss E.H., Weichhart T., Heffeter P, & Weckwerth W. (2024). Targeting PHGDH reverses the immunosuppressive phenotype of tumor-associated macrophages through α-ketoglutarate and mTORC1 signaling. Cellular and Molecular Immunology, 21(5), 448-465.

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Immunoblotting Analyses of Cellular Signaling

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Anti-p70 S6K1 (Santa Cruz Biotechnology, Dallas, TX; SC-230), anti-phospho (T389) p70 S6K1 (Cell Signaling Technology, Danvers, MA; #9205), anti-S6 (Cell Signaling Technology; #2217), anti-phospho (S235/236) S6 (Cell Signaling Technology; #4856), anti-GFP (Santa Cruz Biotechnology; SC-9996), anti-PARP-1 (Santa Cruz Biotechnology; SC-7150), anti-Akt1/2/3 (Santa Cruz Biotechnology; SC-8312), anti-phospho (S473) Akt (Santa Cruz biotechnology; SC-7958), anti-LC3B (Cell Signaling Technology; #2775), anti-p53 (Santa Cruz Biotechnology, Dallas; SC-126), and anti-actin (Millipore, Temecula, CA; mab1501) antibodies were utilized in this study.

Nam K.H., Yi S.A., Nam G., Noh J.S., Park J.W., Lee M.G., Park J.H., Oh H., Lee J., Lee K.R., Park H.J., Lee J, & Han J.W. (2019). Identification of a novel S6K1 inhibitor, rosmarinic acid methyl ester, for treating cisplatin-resistant cervical cancer. BMC Cancer, 19, 773.

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Immunoblotting Analysis of Protein Signaling

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Protein lysates were separated by 11% SDS-PAGE and blotted on Immobilon-P PVDF membranes (Sigma-Aldrich), following the manufacturer’s instructions and using TE 22 Mini Tank transfer unit (GE Healthcare, Waukesha, WI, USA). Membranes were incubated overnight with the following primary antibodies: CK2α/CK2α′ (MCA3031Z; Bio-Rad, Hercules, CA, USA); anti-CK2β (catalog ab76025) from Abcam (Cambridge, UK); anti-p-Akt (Ser473) (catalog #4060), anti-p-PRAS40 (Thr246) (catalog #13175), anti-p-GSK3β (Ser9) (catalog #5558), anti-GSK3β (catalog #9336), anti-p-ERK1/2 (Thr202/Tyr204) (catalog #4370) and anti-ERK1/2 (catalog #4695) from Cell Signaling Technology (Danvers, MA, USA); anti-Akt1/2/3 (catalog sc-8312) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and anti-β-actin (catalog A5441) from Sigma-Aldrich (Burlington, MA, USA). After removal of primary antibodies and washing with Tris-buffered saline (TBS), membranes were incubated with the secondary antibodies towards rabbit and mouse IgG, conjugated to horseradish peroxidase (PerkinElmer). A signal was developed using an enhanced chemiluminescent detection system ECL (Amersham Biosciences, Little Chalfont, UK). Immunostained bands were quantified by means of a Kodak Image Station 4000MM-PRO and analyzed with Carestream Molecular Imaging software (New Haven, CT, USA).

Sanna M., Borgo C., Compagnin C., Favaretto F., Vindigni V., Trento M., Bettini S., Comin A., Belligoli A., Rugge M., Bassetto F., Donella-Deana A., Vettor R., Busetto L, & Milan G. (2020). White Adipose Tissue Expansion in Multiple Symmetric Lipomatosis Is Associated with Upregulation of CK2, AKT and ERK1/2. International Journal of Molecular Sciences, 21(21), 7933.

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Synthesis and Evaluation of NGF-Based Dipeptides

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The dimeric dipeptides GK-6 and GK-2 were synthesized on the base of murine NGF at the Zakusov Institute of Pharmacology (Moscow, Russia).
Inhibitors of PI3K (LY294002) and MAPK (PD98059) were purchased from Tocris Bioscience (Bristol, UK). The tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT),was obtained from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s Modified Eagle’s medium was purchased from HyClone Laboratories (Logan, UT, USA). Fetal bovine serum was obtained from Gibco (Langley, OK, USA). Glutamine was purchased from ICN Pharmaceuticals. Inc. (Costa Mesa, CA, USA). Poly-D-lysine was purchased from BD Biosciences (San Jose, CA, USA). DC protein assay was purchased from Biorad (Hercules, CA, USA). Anti-TrkA, anti-pTrkA, anti-AKT1/2/3, anti-pAKT1/2/3, anti-ERK1/2, anti-pERK1/2 antibodies and enhanced chemiluminescence kits were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-β-actin antibodies and horseradish peroxidase conjugated antibodies were purchased from Abcam (Cambridge, MA, USA).

Gudasheva T.A., Povarnina P.Y., Antipova T.A., Firsova Y.N., Konstantinopolsky M.A, & Seredenin S.B. (2015). Dimeric dipeptide mimetics of the nerve growth factor Loop 4 and Loop 1 activate TRKA with different patterns of intracellular signal transduction. Journal of Biomedical Science, 22, 106.

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PI3-kinase and AKT protein expression

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Each ICB sample was harvested, rinsed with PBS, and lysed. A total of 40 μg of protein per sample was separated by SDS-polyacrylamide gel electrophoresis, incubated with anti-PI3-kinase p85α (Santa Cruz Biotechnology) at a dilution of 1 : 1000 or anti-AKT1/2/3 (Santa Cruz Biotechnology) at a dilution of 1 : 500. Signal was visualized after secondary antibody staining (donkey anti-rabbit IgG antibody, 1 : 4000). All values were normalized with β-actin as loading control. Each sample was collected from two rats (4 eyes).

Zhang N., Yu S., Liu X, & Lu H. (2017). Low Dose of Lipopolysaccharide Pretreatment Preventing Subsequent Endotoxin-Induced Uveitis Is Associated with PI3K/AKT Pathway. Journal of Immunology Research, 2017, 1273940.

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Fluorescent In-Situ Detection of Akt

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The oligonucleotide containing the Zika virus target sequence (bold) (ZTargetamine: 5’-amine-C6-MMT-TAAAGATGGCTGTTGGTATGGAATGGAGATAAGGCCCAGGAAAG-3’) was covalently attached to an IgG secondary antibody (goat anti-rabbit IgG, whole molecule; Sigma), using an oligonucleotide-conjugation kit according to the manufacturers’ procedure (Abcam, Cambridge, MA, USA). HeLa cells were grown in Falcon 8-well cell culture slides, fixed with 10% formalin/PBS, permeabilized with saponin (0.05%) and incubated with primary (anti-Akt1/2/3 (Santa Cruz, Dallas, TX, USA)) and oligonucleotide-labeled secondary antibody using standard protocols as for immunofluorescence staining, and TN-RCA performed in situ essentially as described above, washed with PBS, and photographed with a fluorescent microscope (BZ-X710, Keyence, Itasca, IL, USA).

Zingg J.M, & Daunert S. (2018). Trinucleotide Rolling Circle Amplification: A Novel Method for the Detection of RNA and DNA. Methods and Protocols, 1(2), 15.

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Synthesis and Characterization of Dimeric Dipeptides

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The dimeric dipeptides GSB-106 ((bis-(N-monosuccinyl-l-seryl-l-lysine)hexamethylenediamide (Tm =143°C–145°C, [α] 20 (link) D =−24.7° (c=0.4%; dimethylformamide)) and GSB-214((bis-(N-monosuccinyl-l-methionyl-l-serine) heptamethylenediamide (Tm =160°C–162°C, [α] 20 (link) D =−21.75° (c=0.4%; MeOH)) were synthesized at the Zakusov Institute of Pharmacology (Moscow, Russia), as described previously.13 (link)
2,3,5-triphenyltetrazolium chloride (TTC) and Nembutal were obtained from Sigma-Aldrich (St Louis, MO, USA). Dulbecco’s Modified Eagle’s Medium was purchased from HyClone Laboratories (Logan, UT, USA). Fetal bovine serum was obtained from Gibco (Langley, OK, USA). Glutamine was purchased from ICN Pharmaceuticals, Inc. (Costa Mesa, CA, USA). Poly-d-lysine was purchased from BD Biosciences (San Jose, CA, USA). DC protein assay was purchased from Bio-Rad (Hercules, CA, USA). Anti-TrkA, anti-pTrkA, anti-AKT1/2/3, anti-phospho-AKT1^s473/2^s472/3^s474 anti-ERK1/2, anti-phospho ERK1/2^Y204 antibodies, and enhanced chemiluminescence kits were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-β-actin antibodies and horseradish peroxidase-conjugated antibodies were purchased from Abcam (Cambridge, MA, USA). Formalin was purchased from Ecros (Saint Petersburg, Russia). GSB-106 and GSB-214 were dissolved in water. Then solvents were diluted in culture media in equivalent amounts.

Gudasheva T.A., Povarnina P., Logvinov I.O., Antipova T.A, & Seredenin S.B. (2016). Mimetics of brain-derived neurotrophic factor loops 1 and 4 are active in a model of ischemic stroke in rats. Drug Design, Development and Therapy, 10, 3545-3553.

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Western Blot Protein Analysis

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Samples of the protein fractions containing equivalent amounts of proteins per lane (70 μg per lane) were separated by 10% or 12% SDS-PAGE gel electrophoresis (according to molecular weight of specific protein). For Western blot assays, proteins were transferred to a nitrocellulose membrane. The quality of the transfer was controlled by Ponceau S staining of nitrocellulose membranes after the transfer. Specific anti-Akt 1/2/3 (dilution 1:330), anti-p-Akt 1/2/3 (dilution 1:710), anti-eNOS (dilution 1:200), anti-PKCε (dilution 1:1000), anti-GSK3B (dilution 1:330), anti p-GSK3B (dilution 1:250), anti-BAX (dilution 1:200), anti-Bcl-2 (dilution 1:200) and anti-caspase 3 (dilution 1:100) (Santa Cruz Biotechnology) antibodies were used for the primary immunodetection. Peroxidase-labeled anti-rabbit immunoglobulin (Cell Signaling Technology) a peroxidase-labeled anti-mouse immunoglobulin were used as the secondary antibodies (dilution 1:2000). Bound antibodies were detected by the enhanced chemiluminescence (ECL) method. The optical density of individual bands was analyzed by PCBAS 2.08e software and normalized to GAPDH (anti-GAPDH antibody, dilution 1:750) as an internal control.

Kindernay L., Farkasova V., Neckar J., Hrdlicka J., Ytrehus K, & Ravingerova T. (2021). Impact of Maturation on Myocardial Response to Ischemia and the Effectiveness of Remote Preconditioning in Male Rats. International Journal of Molecular Sciences, 22(20), 11009.

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