Sc 5298

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Sc-5298 is a laboratory instrument designed for the analysis and detection of biological samples. It utilizes advanced spectroscopic techniques to provide precise and reliable data. The core function of Sc-5298 is to facilitate the identification and quantification of specific biomolecules within complex matrices.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Akt (sc-5298) (5 citations) Anti-Akt (sc-5298) (3 citations) Anti-AKT1 sc-5298 (2 citations) AKT1 (sc-5298; (2 citations)

15 protocols using sc 5298

Isoliquiritigenin Platelet Activation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Isoliquiritigenin (961-29-5; the structure is listed in Figure 1A) was purchased from Weikeqi-biotech. Thrombin (T4393), ADP (A5285), PGE₁ (P7527), fibrinogen (F3879), TRITC-Phalloidin (P1951), dimethyl sulfoxide (DMSO; D2650), bovine serum albumin (BSA; B2064), and Triton X-100 (X100) were purchased from Sigma-Aldrich. Collagen (LS001652) was purchased from Chrono-Log. Fluo 3-AM (F023-10) was purchased from Dojindo. Fluorescein-conjugated antibody to human CD62P (Clone: AK-4) and PAC-1 (Clone: PAC-1) were purchased from BD Biosciences. The antibodies for the immunoblot in this study were the following: anti-phospho-PLCγ2 (Tyr759) antibody (50535, CST), anti-phospho-ERK1/2 (Thr202/Tyr204) antibody (4370, CST), anti-ERK1/2 antibody (9194, CST), anti-phospho-Jun N-terminal kinase (JNK) (Thr183/Tyr185) antibody (sc-6254, Santa Cruz), anti-JNK antibody (sc-7345, Santa Cruz), anti-phospho-p38 MAPK (Thr180/Tyr182) antibody (4511, CST), anti-p38 MAPK antibody (8690, CST), anti-phospho-Akt (Ser473) antibody (4060, CST), anti-Akt antibody (sc-5298, Santa Cruz), and anti-GAPDH (ANT011, Antegene).

Wu Y., Deng C., Wang Y., Ming Z., Mei H, & Hu Y. (2023). The inhibitory effect of isoliquiritigenin on human platelets in vitro. Annals of Translational Medicine, 11(6), 250.

+ Open protocol

+ Expand

Immunoprecipitation of Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

For immunoprecipitation assays, cell lysates were incubated with specific antibodies against MEK1 (Santa Cruz Biotechnology, sc-6250, 1:100), ERK1/2 (Santa Cruz Biotechnology, sc-514302, 1:100), AKT1 (Santa Cruz Biotechnology, sc-5298, 1:100) or DRP1 (Santa Cruz Biotechnology, sc-101270, 1:100) at 4°C overnight. After addition of Gammabind G Sepharose beads, samples were incubated at 4°C for 4 h. Beads were washed three times with cold PBS and proteins were released from the beads by boiling in SDS-sample loading buffer and analyzed by SDS-PAGE.

Cha Y., Kim T., Jeon J., Jang Y., Kim P.B., Lopes C., Leblanc P., Cohen B.M, & Kim K.S. (2021). SIRT2 regulates mitochondrial dynamics and reprogramming via MEK1-ERK-DRP1 and AKT1-DRP1 axes. Cell reports, 37(13), 110155.

+ Open protocol

+ Expand

Western Blot Analysis of Apoptotic Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot analysis was performed on renal lysates or cell extracts using the following primary antibodies: anti-DcR2 (ab108421; Abcam), anti-FLIP (ab8421; Abcam), anti-cleaved caspase 3 (ab214430; Abcam), anti-caspase 8 (ab25901; Abcam), caspase 3 (ab184787; Abcam), anti-caspase 7 (ab255818; Abcam), anti-cleaved caspase 7 (ab256469; Abcam), Akt (sc5298, Santa Cruz), pAkt (sc135650, Santa Cruz), and anti-GAPDH (BM3876; Boster). The intensity of each band was analyzed using Quantity One software (Bio-Rad, Hercules, CA, USA).

Chen J., Chen K.H., Wang L.M., Luo J., Zheng Q.Y, & He Y.N. (2022). Decoy receptor 2 mediates the apoptosis-resistant phenotype of senescent renal tubular cells and accelerates renal fibrosis in diabetic nephropathy. Cell Death & Disease, 13(6), 522.

+ Open protocol

+ Expand

Comprehensive Antibody and Inhibitor Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies against VEGF (SC-507), focal adhesion kinase (FAK; SC-932), Src (SC-5226), Akt (SC-5298), and CD34(SC-74499) were bought from Santa Cruz (Santa Cruz, CA, USA). Antibodies targeting p-FAK (3283S), p-Src (2101S), p-AKT (4060S), and CD133(64326s) were obtained from Cell Signaling Technology, Inc. (Beverly, MA, USA). Antibodies against CD31(ab28364) was purchased from Abcam (Cambridge, MA, USA). Small interfering RNAs (siRNAs) against FAK (L-003164-00), Src (L-003110-00) and Akt (L-003000-00-0005) and their respective controls were purchased from Dharmacon (Lafayette, CO, USA). We purchased VEGF shRNA plasmids from the National RNAi Core (Taipei, Taiwan). Inhibitors for FAK (869288-64-2) were from Calbiochem (San Diego, CA, USA). A VEGF ELISA kit (DY293B) was purchased from R&D Systems (Minneapolis, MN, USA) and APLN ELISA (KA1681) kit was purchased from Abnova (Taipei, Taiwan). Inhibitors for Src (P0042), Akt (A6730) and all the chemicals not mentioned above were supplied by Sigma-Aldrich (St. Louis, MO, USA).

Wang Y.H., Kuo S.J., Liu S.C., Wang S.W., Tsai C.H., Fong Y.C, & Tang C.H. (2020). Apelin Affects the Progression of Osteoarthritis by Regulating VEGF-Dependent Angiogenesis and miR-150-5p Expression in Human Synovial Fibroblasts. Cells, 9(3), 594.

+ Open protocol

+ Expand

FCL Inhibits Cancer Cell Migration

Check if the same lab product or an alternative is used in the 5 most similar protocols

FCL (purity >98%) was purchased from Shanghai Standard Technology Co., Ltd. (Shanghai, China). Cell culture materials were purchased from Gibco (Thermo Fisher Scientific, Inc., Waltham, MA, USA). Specific antibodies against GAPDH (1:1,000, sc-25778), MMP-2 (1:500, sc-53630), MMP-9 (1:500, sc-21733), phosphorylated (p)-AKT (1:500, sc-135650) and AKT (1:500, sc-5298) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Chen Z., He T., Zhao K, & Xing C. (2016). Anti-metastatic activity of fangchinoline in human gastric cancer AGS cells. Oncology Letters, 13(2), 655-660.

+ Open protocol

+ Expand

Murine Lung Protein Analysis Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The murine lung tissue samples were made into homogenate and centrifuged at 20,000 at 4°C for 30 min to collect total protein, and the protein concentration was determined using the bicinchoninic acid method (23225, Pierce Biotechnology, Waltham, MA, USA). Then, the protein sample was separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto polyvinylidene fluoride membranes. The membranes were incubated with the primary antibodies against SP1 (sc-420, 1:1,500, Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA), PTEN (MA5-12,278, 1:2,000, Thermo Fisher Scientific), AKT (sc-5298, 1:1,500, Santa Cruz), p-AKT (NB100-56749, 1:1,000, Novus Biologicals, Littleton, CO, USA) and GAPDH (ab8245, 1:2,000, Abcam) at 4°C for 16 h, and then with the secondary antibody (ab205719, 1:5,000, Abcam) at 37°C for 3 h. The protein bands were analyzed using an enhanced chemiluminescence (ECL) kit (PE0010, Solarbio) [31 (link)].

Zhao H., Shi L., Wang X., Yu X, & Wang D. (2022). Sp1 transcription factor represses transcription of phosphatase and tensin homolog to aggravate lung injury in mice with type 2 diabetes mellitus-pulmonary tuberculosis. Bioengineered, 13(4), 9928-9944.

+ Open protocol

+ Expand

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

RIPA buffer (pH = 8.0) including 150 mM NaCl, 0.1% SDS, 50 mM Tris-HCl and 1% NP-40 was added to the proteinase inhibitor (Roche Diagnostics, Co., Ltd., Rotkreuz, Switzerland) and used to obtain whole cell lysates. The proteins were examined using a BCA kit (Thermo Fisher) and separated by SDS-PAGE. The proteins were transferred onto PVDF membranes (0.45 µm, Millipore Corp, Billerica, MA, USA), sealed with 5% BSA for 60 min at 25°C, and hybridized at 4°C with primary antibodies to MMP2 (1:1000, MA5-14,186, Invitrogen Inc., Carlsbad, CA, USA), MMP9 (1:2000, ab58803, Abcam), Bax (1:1200, sc-7480, Santa Cruz Biotechnology), PI3K (1:1,800, sc-8010, Santa Cruz Biotechnology), p-AKT (1:1,500, ab81283, Abcam), AKT (1:1,300, sc-5298, Santa Cruz Biotechnology), and GAPDH (1:3,000, G8795, Sigma-Aldrich, St Louis, MO, USA) for one night. The blots were incubated with secondary antibodies for 60 min at 25°C. Finally, the protein bands were detected using an ECL kit (Thermo Fisher Scientific). Densitometric analysis was performed using IPP 6.0 (Image-Pro Plus 6.0).

Li P., Lv H., Wu Y., Xu K., Xu M, & Ma Y. (2021). E2F transcription factor 1 is involved in the phenotypic modulation of esophageal squamous cell carcinoma cells via microRNA-375. Bioengineered, 12(2), 10047-10062.

+ Open protocol

+ Expand

Correlation of HK2, Akt1, p-Akt1, and FN1 in SCC

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human squamous cervical carcinoma (SCC) samples used in this study were collected at the First Affiliated Hospital of Xi’an Jiaotong University from 2008 to 2016 as described in our previous study. The immunohistochemical staining procedure and the evaluation standard of immunohistochemistry (IHC) score were performed as previously described [18 (link)]. To validate the correlation between HK2 and Akt1, p-Akt1, FN1 expression in vivo, serial sections of human squamous cervical carcinoma samples (n = 15) were immunostained with an anti-HK2 (1:100 dilution, sc-374091, Santa Cruz, USA), anti-Akt1 (1:100 dilution, sc-5298, Santa Cruz, USA), anti-p-Akt1 (1:100 dilution, sc-293125, Santa Cruz, USA), anti-FN1 (1:300 dilution, 15613-1-AP, Wuhan, China). The immunohistochemistry (IHC) score of HK2, Akt1, p-Akt1 and FN1 in these SCC samples was confirmed by using Pearson correlation analysis.

Chen Q., Li L., Liu X., Feng Q., Zhang Y., Zheng P, & Cui N. (2021). Hexokinases 2 promoted cell motility and distant metastasis by elevating fibronectin through Akt1/p-Akt1 in cervical cancer cells. Cancer Cell International, 21, 600.

+ Open protocol

+ Expand

Western Blot Analysis of Cell Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were prepared with lysis buffer (1% Triton X-100, 150 mmol/L NaCl, 10 mmol/L Tris-HCl, pH 7.4, 1 mmol/L EDTA, 1 mmol/L EGTA, pH 8.0, 0.2 mmol/L Na₃VO₄, 0.2 mmol/L phenylmethylsulfonyl fluoride (PMSF), and 0.5% nonidet P (NP)-40). Equal amounts of protein (60–100 μg) were separated by 10% SDS-PAGE and electro-transferred on to a polyvinylidenefluoride (PVDF) membrane. Membranes were blocked with 5% non-fat milk in tris-buffered saline-Tween (TBST) for 2 h at room temperature and incubated with specific primary antibodies against cyclin E1 (1:500, sc-377,100, Santa Cruz), CDK1 (1: 500, ab265590, Abcam), PIK3CA (1:500, sc-293,172, Santa Cruz), AKT1 (1:500, sc-5298, Santa Cruz), p-AKT (1:500, #4060, Cell Signaling Technology), cleaved-caspase 8 (1:500, #8592P, Cell Signaling Technology), or tubulin (1:1000, ab7291, Abcam) at 4 °C overnight. The membranes were then incubated with HRP-conjugated secondary antibody (1:1000, Seracare) for 1 h at room temperature, following the blots were visualized using GE ImageQuant™ LAS 4000 detection system. The protein bands of interest were quantified using Quantity One software (Bio-Rad).

Zhang Y., Lv P., Ma J., Chen N., Guo H., Chen Y., Gan X., Wang R., Liu X., Fan S., Cong B, & Kang W. (2021). Antrodia cinnamomea exerts an anti-hepatoma effect by targeting PI3K/AKT-mediated cell cycle progression in vitro and in vivo. Acta Pharmaceutica Sinica. B, 12(2), 890-906.

+ Open protocol

+ Expand

Immunoprecipitation of Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!