Anti phospho aktser473

Manufactured by Bioworld Technology

Sourced in United States

Anti-phospho-AktSer473 is a laboratory reagent used to detect the phosphorylation of the serine 473 residue of the Akt protein. This antibody is commonly used in research applications to study the activation and regulation of the Akt signaling pathway.

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Lab products found in correlation

Western bright ecl detection system, by Advansta (4 mentions) Anti gapdh, by Abcepta (4 mentions) Pvdf membrane, by Roche (4 mentions) Anti ampk thr172, by Cell Signaling Technology (4 mentions) Uric acid, by Merck Group (4 mentions) N 7 nitrobenz 2 oxa 1 3diazol 4 yl amino 2 deoxy d glucose 2 nbdg, by Thermo Fisher Scientific (4 mentions) Anti phospho irs1 ser307, by Thermo Fisher Scientific (4 mentions) 5 amino 4imidazole 1 β d carboxamide ribofuranoside aicar, by MedChemExpress (4 mentions) Anti akt ser473, by Bioworld Technology (4 mentions) Compound c, by Merck Group (4 mentions) Insulin, by Merck Group (4 mentions) Metformin, by Merck Group (4 mentions) Anti phospho erk1 2, by Bioworld Technology (3 mentions) Anti total akt, by Bioworld Technology (3 mentions) Species specific hrp conjugated secondary antibodies, by ZSGB-BIO (3 mentions) Anti phospho ampk thr172, by Cell Signaling Technology (3 mentions) Rabbit anti gapdh antibody, by Abcam (3 mentions) Anti irs1 ser307, by Thermo Fisher Scientific (3 mentions) Anti total erk, by Bioworld Technology (2 mentions) Anti ehf, by Abcam (2 mentions)

8 protocols using anti phospho aktser473

Phospho-protein Detection Protocol

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The indicated cells were lysed in RIPA buffer containing protease inhibitors and phosphatase inhibitor Cocktail (Sigma, Mo, USA). Supernatants were collected and subjected to 10% SDS-PAGE, and transferred onto PVDF membranes (Roche Diagnostics, Mannheim, Germany). The membranes were then incubated overnight with primary antibodies. Anti-phospho-Akt^Ser473, anti-phospho-Erk1/2, anti-total Akt and anti-total Erk were purchased from Bioworld Technology, co, Ltd. Anti-EHF was purchased from Abcam Biotechnology, Inc. Anti-GAPDH was purchased from Abgent, Inc. This was followed by incubation with species-specific HRP-conjugated secondary antibodies from ZSGB-BIO, and antigen-antibody complexes were visualized using the Western Bright ECL detection system (Advansta, CA).

Lv Y., Sui F., Ma J., Ren X., Yang Q., Zhang Y., Guan H., Shi B., Hou P, & Ji M. (2016). Increased expression of EHF contributes to thyroid tumorigenesis through transcriptionally regulating HER2 and HER3. Oncotarget, 7(36), 57978-57990.

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Protein Expression Analysis by Western Blot

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Cells were lysed in prechilled RIPA buffer containing protease inhibitors. Equal amounts of protein lysates were separated by SDS–PAGE and transferred onto PVDF membranes (Roche Diagnostics, Mannheim, Germany). The membranes were then incubated with primary antibodies. Anti-AIB1 and anti-total-Erk1/2 (t-Erk) were purchased from Abcam, Inc. Anti-phospho-Akt^Ser473, anti-phospho-Erk1/2 and anti-total-Akt (t-Akt) were purchased from Bioworld Technology, co, Ltd. Anti-total-Rb (t-Rb) and anti-phospho-Rb^S811 (p-Rb) were purchased from Epitomics, Inc. Anti-GAPDH was purchased from Abgent, Inc. This was followed by incubation with species-specific HRP-conjugated secondary antibodies from ZSGB-BIO, and immunoblotting signals were visualized using the Western Bright ECL detection system (Advansta, CA).

Shi J., Liu W., Sui F., Lu R., He Q., Yang Q., Lv H., Shi B, & Hou P. (2015). Frequent amplification of AIB1, a critical oncogene modulating major signaling pathways, is associated with poor survival in gastric cancer. Oncotarget, 6(16), 14344-14359.

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Western Blot Analyses of Signaling Proteins

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Cells were lysed in prechilled RIPA buffer containing protease inhibitors. The protein lysates were separated on SDS–PAGE and then transferred to PVDF membranes (Roche Diagnostics, Mannheim, Germany). The membranes were blocked for 2 h in 5% bovine serum albumin (BSA) in 1 × TBS-T (0.5% Tween-20) and incubated with the indicated primary antibodies, including anti-EHF (Abcam, Inc), anti-total-Erk1/2 (Abcam, Inc), anti-phospho-Erk1/2 (Epitomics, Inc), anti-phospho-AktSer473 (Bioworld Technology, co, Ltd), anti-total-Akt (Bioworld Technology, co, Ltd), anti-HER2 (Sino Biological, Inc), anti-HER3 (Sino Biological, Inc), anti-HER4 (Sino Biological, Inc), anti-E-cadherin (Epitomics, Inc), anti-Vimentin (Epitomics, Inc) and anti-GAPDH (Abgent, Inc). The membranes were then incubated with species-specific HRP-conjugated secondary antibodies from ZSGB-BIO, and immunoblotting signals were visualized using the Western Bright ECL detection system (Advansta, CA).

Shi J., Qu Y., Li X., Sui F., Yao D., Yang Q., Shi B., Ji M, & Hou P. (2016). Increased expression of EHF via gene amplification contributes to the activation of HER family signaling and associates with poor survival in gastric cancer. Cell Death & Disease, 7(10), e2442-.

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Immunoblotting Analysis of Cellular Signaling

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The indicated cells were lysed in RIPA buffer containing protease inhibitors. Supernatants were collected and subjected to 10% SDS-PAGE, and transferred onto PVDF membranes (Roche Diagnostics, Mannheim, Germany). The membranes were then incubated overnight with primary antibodies. Anti-N-cadherin was purchased from Abcam Biotechnology, Inc. Anti-phospho-Erk1/2, anti-phospho-Akt^Ser473, anti-total Erk and anti-total Akt were purchased from Bioworld Technology, co, Ltd. Anti-p16, anti-phospho-Rb and anti-Rb antibodies were purchased from Abcam. Anti-GAPDH was purchased from Abgent, Inc. This was followed by incubation with species-specific HRP-conjugated secondary antibodies from ZSGB-BIO, and antigen-antibody complexes were visualized using the Western Bright ECL detection system (Advansta, CA).

Da C., Wu K., Yue C., Bai P., Wang R., Wang G., Zhao M., Lv Y, & Hou P. (2016). N-cadherin promotes thyroid tumorigenesis through modulating major signaling pathways. Oncotarget, 8(5), 8131-8142.

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Insulin Signaling Pathway Assay

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Uric acid (15 mg/dL, final concentration), metformin, insulin (100 nM, final concentration) and compound C (20 μM, final concentration) were purchased from Sigma. N‐(7‐Nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl)amino]‐2‐deoxy‐d‐glucose (2‐NBDG, 100 μM, final concentration), Alexa Fluor®–conjugated secondary antibodies, anti‐IRS1 and anti‐phospho‐IRS1 (Ser307) antibodies were purchased from Invitrogen. 5‐Amino‐4‐imidazole‐1‐β‐D‐carboxamide ribofuranoside (AICAR, 500 μM, final concentration) was purchased from MedChemExpress (MCE LLC, USA). A rabbit anti‐GAPDH and anti‐GLUT4 antibody were purchased from Abcam (Abcam, USA). Anti‐Akt (Ser473) and anti‐phospho‐Akt (Ser473) antibodies were purchased from Bioworld (St. Louis Park, USA). Anti‐AMPK (Thr172) and anti‐phospho‐AMPK (Thr172) antibodies were purchased from Cell Signaling Technology (CST, Beverly). Cat no for antibodies: anti‐phospho‐IRS1 (Ser307) (44‐813G); anti‐phospho‐AMPK (Thr172) (50081); anti‐phospho‐Akt (Ser473) (APG01691G); anti‐GLUT4 (ab33780).

Jiao Z., Chen Y., Xie Y., Li Y, & Li Z. (2021). Metformin protects against insulin resistance induced by high uric acid in cardiomyocytes via AMPK signalling pathways in vitro and in vivo. Journal of Cellular and Molecular Medicine, 25(14), 6733-6745.

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Metabolic Regulation in Cardiomyocytes

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Uric acid (15 mg/dl, final concentration), metformin, insulin (100 nM, final concentration) and compound C (20 μM, final concentration) were purchased from Sigma (St. Louis, MO, USA). N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG, 100 μM, final concentration) and anti-IRS1 (Ser307) and anti-phospho-IRS1 (Ser307) antibodies were purchased from Invitrogen (Carlsbad, CA, USA). 5-Amino-4-imidazole-1-β-D-carboxamide ribofuranoside (AICAR, 500 μM, final concentration) was purchased from MedChemExpress (MCE LLC, USA). A rabbit anti-GAPDH antibody was purchased from Abcam (Abcam, USA). Anti-Akt (Ser473) and anti-phospho-Akt (Ser473) antibodies were purchased from Bioworld (St. Louis Park, USA). Anti-AMPK (Thr172) and anti-phospho-AMPK (Thr172) antibodies were purchased from Cell Signaling Technology (CST, Beverly, MA, USA). Cardiomyocytes were cultured in serum-free media for 24 hours and then incubated in the presence of 15 mg/dl HUA for 24 hours. Cardiomyocytes were pretreated with either metformin (0 to 20 μmol/L) or AICAR, an AMPK activator (500 μmol/L), for 60 minutes before the addition of HUA. Other cells were preincubated with compound C for 6 hours before the addition of either metformin or AICAR. Then, 2-NBDG uptake was analyzed in cardiomyocytes. were from Millipore (Billerica, MA).

Jiao Z., Chen Y., Xie Y., Li Y., & Li Z. (2021). Metformin protects against insulin resistance induced by high uric acid in cardiomyocytes via AMPK signaling pathways in vitro and in vivo.

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Metabolic Regulation in Cardiomyocytes

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Uric acid (15 mg/dl, nal concentration), metformin, insulin (100 nM, nal concentration) and compound C (20 µM, nal concentration) were purchased from Sigma (St. Louis, MO, USA). N-(7-Nitrobenz-2-oxa-1,3diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG, 100 µM, nal concentration) and anti-IRS1 (Ser307) and anti-phospho-IRS1 (Ser307) antibodies were purchased from Invitrogen (Carlsbad, CA, USA). 5-Amino-4imidazole-1-β-D-carboxamide ribofuranoside (AICAR, 500 µM, nal concentration) was purchased from MedChemExpress (MCE LLC, USA). A rabbit anti-GAPDH antibody was purchased from Abcam (Abcam, USA). Anti-Akt (Ser473) and anti-phospho-Akt (Ser473) antibodies were purchased from Bioworld (St. Louis Park, USA). Anti-AMPK (Thr172) and anti-phospho-AMPK (Thr172) antibodies were purchased from Cell Signaling Technology (CST, Beverly, MA, USA). Cardiomyocytes were cultured in serum-free media for 24 hours and then incubated in the presence of 15 mg/dl HUA for 24 hours. Cardiomyocytes were pretreated with either metformin (0 to 20 µmol/L) or AICAR, an AMPK activator (500 µmol/L), for 60 minutes before the addition of HUA. Other cells were preincubated with compound C for 6 hours before the addition of either metformin or AICAR. Then, 2-NBDG uptake was analyzed in cardiomyocytes.
were from Millipore (Billerica, MA).

Jiao Z., Chen Y., Xie Y., Li Y., & Li Z. (2021). Metformin protects against insulin resistance induced by high uric acid in cardiomyocytes via AMPK signaling pathways in vitro and in vivo.

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Glucose Uptake in Cardiomyocytes

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Uric acid (15 mg/dl, nal concentration), metformin, insulin (100 nM, nal concentration) and compound C (20 µM, nal concentration) were purchased from Sigma (St. Louis, MO, USA). N-(7-Nitrobenz-2-oxa-1,3diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG, 100 µM, nal concentration) and anti-IRS1 (Ser307) and anti-phospho-IRS1 (Ser307) antibodies were purchased from Invitrogen (Carlsbad, CA, USA). 5-Amino-4imidazole-1-β-D-carboxamide ribofuranoside (AICAR, 500 µM, nal concentration) was purchased from MedChemExpress (MCE LLC, USA). A rabbit anti-GAPDH antibody was purchased from Abcam (Abcam, USA). Anti-Akt (Ser473) and anti-phospho-Akt (Ser473) antibodies were purchased from Bioworld (St.
Louis Park, USA). Anti-AMPK (Thr172) and anti-phospho-AMPK (Thr172) antibodies were purchased from Cell Signaling Technology (CST, Beverly, MA, USA). Cardiomyocytes were cultured in serum-free media for 24 hours and then incubated in the presence of 15 mg/dl HUA for 24 hours. Cardiomyocytes were pretreated with either metformin (0 to 20 µmol/L) or AICAR, an AMPK activator (500 µmol/L), for 60 minutes before the addition of HUA. Other cells were preincubated with compound C for 6 hours before the addition of either metformin or AICAR. Then, 2-NBDG uptake was analyzed in cardiomyocytes.
were from Millipore (Billerica, MA).

Li Z., Jiao Z., Chen Y., Xie Y., & Li Y. (2021). Metformin Protects Against Insulin Resistance Induced by High Uric Acid in Cardiomyocytes via AMPK Signaling Pathways in Vitro and in Vivo.

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