Log In Sign up
The largest database of trusted experimental protocols

Phosphor akt s473

Manufactured by Cell Signaling Technology
Visit Supplier
Sourced in United States

Phosphor-Akt S473 is a lab equipment product that detects phosphorylation of Akt at serine 473. It is used to measure the activation status of the Akt signaling pathway.

Automatically generated - may contain errors

Lab products found in correlation

α tubulin, by Merck Group (3 mentions) β catenin, by Santa Cruz Biotechnology (3 mentions) Phospho erk1 2, by Cell Signaling Technology (3 mentions) Phospho stat3, by Cell Signaling Technology (3 mentions) Cleaved caspase 3, by Cell Signaling Technology (2 mentions) Quantity one software, by Bio-Rad (2 mentions) Phospho s6 ribosomal protein, by Cell Signaling Technology (2 mentions) Cyclin d1, by Cell Signaling Technology (2 mentions) Phospho irs1 s636 639, by Cell Signaling Technology (2 mentions) Phosphor 4e bp1, by Cell Signaling Technology (2 mentions) Clarity western ecl substrate, by Bio-Rad (2 mentions) Igf 1r, by Cell Signaling Technology (2 mentions) Gapdh, by Santa Cruz Biotechnology (2 mentions) T8203, by Merck Group (2 mentions) Mca497rt, by Abcam (2 mentions) Ab14041, by Merck Group (2 mentions) Ab8021, by Abcam (2 mentions) Ab5076, by Bio-Rad (2 mentions) Ab39626, by Santa Cruz Biotechnology (2 mentions) Ab32144, by Abcam (2 mentions)

Spelling variants of this product

Phosphor-AKT (75 citations)

8 protocols using phosphor akt s473

1

Immunoblotting Analysis of Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in NP40 buffer and analyzed by standard immunoblotting protocols. Antibodies used were directed against: ERK1/2, phospho-ERK1/2, phospho-STAT3, phosphor-Akt S473, IRS1, phospho-IRS1 S636/639, IGF-1R, phospho-S6 ribosomal protein, phosphor-4E-BP1, cyclin D1, cleaved caspase 3 (Cell Signaling, Danvers, MA, USA); β-catenin, SHP2, IRS1, GAPDH (Santa Cruz Biotechnology, Dallas, TX, USA); and α-tubulin (Sigma-Aldrich). Secondary antibodies were from Cell Signaling. Detection was done using Clarity™ Western ECL Substrate (Biorad, Hercules, CA, USA). Densitometric analysis was perfomed using Quantity One Software (Biorad).
Sanchez-Lopez E., Flashner-Abramson E., Shalapour S., Zhong Z., Taniguchi K., Levitzki A, & Karin M. (2015). Targeting colorectal cancer via its microenvironment by inhibiting IGF-1 Receptor-insulin receptor substrate and STAT3 signaling. Oncogene, 35(20), 2634-2644.
+ Open protocol
+ Expand
2

Immunohistochemical and ELISA Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fluorescent-labelled antibodies for flow cytometry were from eBioscience. Immunoblot analysis and immunohistochemistry were performed with antibodies to Ki-67, MUC2 (GeneTex), active caspase 3, ERK1/2, phospho-ERK1/2, phospho-p38, phospho-JNK, phospho-NF-κB p65, phospho-STAT3, phosphor-Akt S473, phosphor-Akt T308 (Cell Signaling), β-catenin, HDAC1, NF-κB p65, p38 (Santa Cruz Biotechnology), CD11b, gp38, Lyve1, EpCam (eBioscience), CD31 (BD), α-smooth muscle actin (Abcam) and α-tubulin (Sigma). Secondary antibodies (host: donkey) for fluorescent microscopy labelled with Alexa 488 or 594 were from Life Technology. For ELISA analysis of IL-6, colonic tumors were cultured in DMEM containing 10% FBS for overnight and supernatant was analyzed by IL-6 ELISA kit from eBioscience. Cytokine concentration was normalized to the weight of tumors in each well.
Wang K., Kim M.K., Di Caro G., Wong J., Shalapour S., Wan J., Zhang W., Zhong Z., Sanchez-Lopez E., Wu L.W., Taniguchi K., Feng Y., Fearon E., Grivennikov S.I, & Karin M. (2014). Interleukin-17 receptor A signaling in transformed enterocytes promotes early colorectal tumorigenesis. Immunity, 41(6), 1052-1063.
+ Open protocol
+ Expand
3

ARF6 activation in infected cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
GTP-bound ARF6 in infected monolayers was measured per the manufacturer’s instructions (Cytoskeleton, Inc.). Briefly, cells were infected at an MOI of 300:1 (as described above), followed by incubation for an additional 10 or 30 min, lysis on ice, and harvesting and snap-freezing of the lysate. Two hundred fifty micrograms of each lysate was incubated with beads covalently conjugated to the ARF6 binding domain of GGA3 for 1 h at 4°C. Beads were washed and recovered. Bound ARF6 was resolved by SDS-PAGE and analyzed using ARF6 antibody (gift of J. Donaldson). Other antibodies used were phosphor-Akt S473 and pan-Akt (Cell Signaling) and ARNO (Abcam ab56510).
Garza-Mayers A.C., Miller K.A., Russo B.C., Nagda D.V, & Goldberg M.B. (2015). Shigella flexneri Regulation of ARF6 Activation during Bacterial Entry via an IpgD-Mediated Positive Feedback Loop. mBio, 6(2), e02584-14.
+ Open protocol
+ Expand
4

Immunoblotting Analysis of Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in NP40 buffer and analyzed by standard immunoblotting protocols. Antibodies used were directed against: ERK1/2, phospho-ERK1/2, phospho-STAT3, phosphor-Akt S473, IRS1, phospho-IRS1 S636/639, IGF-1R, phospho-S6 ribosomal protein, phosphor-4E-BP1, cyclin D1, cleaved caspase 3 (Cell Signaling, Danvers, MA, USA); β-catenin, SHP2, IRS1, GAPDH (Santa Cruz Biotechnology, Dallas, TX, USA); and α-tubulin (Sigma-Aldrich). Secondary antibodies were from Cell Signaling. Detection was done using Clarity™ Western ECL Substrate (Biorad, Hercules, CA, USA). Densitometric analysis was perfomed using Quantity One Software (Biorad).
Sanchez-Lopez E., Flashner-Abramson E., Shalapour S., Zhong Z., Taniguchi K., Levitzki A, & Karin M. (2015). Targeting colorectal cancer via its microenvironment by inhibiting IGF-1 Receptor-insulin receptor substrate and STAT3 signaling. Oncogene, 35(20), 2634-2644.
+ Open protocol
+ Expand
5

Evaluation of IGF-1R Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols
Eca-109 and HEK-293T cells were cultured in DMEM and TE-1 cells were cultured in RPMI-1640 medium, supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis) at 37°C and 5% CO2. Anti-IGF-1R(H60), anti-GRK2(C15), anti-GRK6(C-20), anti-ubiquitin(P4D1), β-arrestin-1(K16) and anti-GAPDH(FL-335)antibodies were purchased from Santa Cruz Biotechnology Inc.(Santa Cruz, CA, USA). Antibodies for IGF-1R, phosphor-IGF-1R (Tyr 1135), phosphor-ERK1/2(Thr202/Tyr204) and phosphor-Akt (S473) were from Cell Signaling Technology (Danvers, USA). Antibodies against β-arrestin-1 was purchased from abcam(Cambridge, MA). IGF-1was from Life Technologies. The other reagents were from Beyotime (Jiangsu, China).
Zhang T., Shen H., Dong W., Qu X., Liu Q, & Du J. (2014). Antitumor effects and molecular mechanisms of figitumumab, a humanized monoclonal antibody to IGF-1 receptor, in esophageal carcinoma. Scientific Reports, 4, 6855.
+ Open protocol
+ Expand
6

Gastrocnemius Muscle Ischemia Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Gastrocnemius muscle from the left (non-ischemic) and right (ischemic) hindlimbs was excised on day 21 after surgery, flash frozen in liquid nitrogen and stored at −80°C until ready for tissue homogenization and processing. The following antibodies were purchased from Cell Signaling Technology, Danvers, MA: PAK1 (#2602), PAK2 (#2608), pPAK2 (Ser20) (#2607), PAK3 (#2609), pan AKT (#4691) phospho-ERK1/2 (Thr202/Tyr204) (#4377), phosphor-AKT (S473) (#9271) and (T308) (#4056). GAPDH antibody (sc-25778) was purchased from Santa Cruz Biotechnologies, Santa Cruz, CA. HRP-conjugated secondary antibodies were purchased from GE Healthcare, Waukesha, WI.
Elsherif L., Ozler M., Zayed M.A., Shen J.H., Chernoff J., Faber J.E, & Parise L.V. (2014). Potential Compensation among Group I PAK Members in Hindlimb Ischemia and Wound Healing. PLoS ONE, 9(11), e112239.
+ Open protocol
+ Expand
7

Immunofluorescent and Immunoblot Analyses of GBM Tumor Markers

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunoblotting and immunofluorescent staining were performed as described65 (link), 66 (link). Specific antibodies against POSTN (Abcam, ab14041, 1: 400), GSC marker SOX2 (Millipore, AB5603, 1:200; Santa Cruz, sc-17320, 1:200) or OLIG2 (R&D systems, AF2418, 1:100), endothelial cell marker Glut1 (Millipore, 07-1401, 1:500) or CD31 (Dako, M082301, 1:100), and macrophage markers Iba1 (Abcam, ab5076, 1:200), CD11b (Abd serotec, MCA711GT, 1:100), Fizz1 (Abcam, ab39626, 1:100), CD163 (Santa Cruz, sc-33560, 1:100), F4/80 (Abd serotec, MCA497RT, 1:100), CX3CR1 (Abcam, ab8021, 1:100) or CCR2 (Abcam, ab32144, 1:100) were used for immunofluorescent staining on GBM tumor sections as indicated. Specific antibodies against POSTN (Abcam, ab14041, 1: 10,000), tubulin (Sigma, T8203, 1:10,000), phosphor-S473 Akt (Cell Signaling Technology, 9271, 1:1,000), Akt (Cell Signaling Technology, 9272, 1:1,000) were used for immunoblot analyses. To determine TAM density, cryosections of GBM tumors were co-stained with Iba1 (or CD11b) and Glut1. The numbers of Iba1+ (or CD11b+) cells were calculated by ImageJ. Glut1 positive areas were regarded as vessels and determined by ImageJ. For TAMs density in each sample, the number of Iba1+ (or CD11+) cells was divided by vessel area for normalization to exclude the individual variation in vascularization. The final outcome was defined as the TAM density.
Zhou W., Ke S.Q., Huang Z., Flavahan W., Fang X., Paul J., Wu L., Sloan A.E., McLendon R.E., Li X., Rich J.N, & Bao S. (2015). Periostin Secreted by Glioblastoma Stem Cells Recruits M2 Tumor-associated Macrophages and Promotes Malignant Growth. Nature cell biology, 17(2), 170-182.
+ Open protocol
+ Expand
8

Immunofluorescent and Immunoblot Analyses of GBM Tumor Markers

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunoblotting and immunofluorescent staining were performed as described65 (link), 66 (link). Specific antibodies against POSTN (Abcam, ab14041, 1: 400), GSC marker SOX2 (Millipore, AB5603, 1:200; Santa Cruz, sc-17320, 1:200) or OLIG2 (R&D systems, AF2418, 1:100), endothelial cell marker Glut1 (Millipore, 07-1401, 1:500) or CD31 (Dako, M082301, 1:100), and macrophage markers Iba1 (Abcam, ab5076, 1:200), CD11b (Abd serotec, MCA711GT, 1:100), Fizz1 (Abcam, ab39626, 1:100), CD163 (Santa Cruz, sc-33560, 1:100), F4/80 (Abd serotec, MCA497RT, 1:100), CX3CR1 (Abcam, ab8021, 1:100) or CCR2 (Abcam, ab32144, 1:100) were used for immunofluorescent staining on GBM tumor sections as indicated. Specific antibodies against POSTN (Abcam, ab14041, 1: 10,000), tubulin (Sigma, T8203, 1:10,000), phosphor-S473 Akt (Cell Signaling Technology, 9271, 1:1,000), Akt (Cell Signaling Technology, 9272, 1:1,000) were used for immunoblot analyses. To determine TAM density, cryosections of GBM tumors were co-stained with Iba1 (or CD11b) and Glut1. The numbers of Iba1+ (or CD11b+) cells were calculated by ImageJ. Glut1 positive areas were regarded as vessels and determined by ImageJ. For TAMs density in each sample, the number of Iba1+ (or CD11+) cells was divided by vessel area for normalization to exclude the individual variation in vascularization. The final outcome was defined as the TAM density.
Zhou W., Ke S.Q., Huang Z., Flavahan W., Fang X., Paul J., Wu L., Sloan A.E., McLendon R.E., Li X., Rich J.N, & Bao S. (2015). Periostin Secreted by Glioblastoma Stem Cells Recruits M2 Tumor-associated Macrophages and Promotes Malignant Growth. Nature cell biology, 17(2), 170-182.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!