Log In Sign up
The largest database of trusted experimental protocols

P akt s473 4060

Manufactured by Cell Signaling Technology
Visit Supplier

The P‐AKT S473 #4060 is an antibody product developed by Cell Signaling Technology. It is designed to detect levels of AKT phosphorylated at serine 473 in various sample types.

Automatically generated - may contain errors

Lab products found in correlation

Chemidoc imaging system, by Bio-Rad (2 mentions) Vegfr2 9698, by Cell Signaling Technology (2 mentions) P plcγ2 4511, by Cell Signaling Technology (2 mentions) Vegfr1 antibody, by R&D Systems (2 mentions) Perkt202 y204 4370, by Cell Signaling Technology (2 mentions) Pfrs2y436 ab193363, by Abcam (2 mentions) Antifade solution, by Thermo Fisher Scientific (1 mentions) Cleaved caspase 3 9664, by Cell Signaling Technology (1 mentions) Lsm 780 laser scanning confocal microscope, by Zeiss (1 mentions) Alexa fluorophore conjugated rabbit or mouse secondary antibodies, by Thermo Fisher Scientific (1 mentions) Rapamycin, by Selleck Chemicals (1 mentions) Gemcitabine, by Selleck Chemicals (1 mentions) γh2ax 05 636, by Merck Group (1 mentions) Pp242, by Selleck Chemicals (1 mentions)

Spelling variants of this product

P-AKT (2452 citations) Akt/p-Akt (17 citations)

6 protocols using p akt s473 4060

1

Immunofluorescence Staining of Cell Cultures

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cultures were fixed with 4% paraformaldehyde/PBS, permeabilized with 0.5% Triton X-100/PBS, and washed 3 times with PBS buffer (137 mM NaCl, 2.7 mM KCl, 4.3 mM Na2HPO4, 1.47 mM KH2PO4, Adjust to a final pH of 7.4). Primary antibodies were suspended in IF buffer (3% bovine serum albumin, 0.1% NP-40 in PBS) and incubated overnight. The cultures were washed with 0.1% NP-40/PBS 6 times and incubated with Alexa fluorophore conjugated rabbit or mouse secondary antibodies (Life Technologies) in IF buffer. The slides were washed with 0.1% NP-40/PBS 6 times, counterstained with 4', 6-diamidino-2-phenylindole (DAPI), and mounted with antifade solution (Life Technologies). Confocal microscopy was done using the Zeiss LSM 780 laser scanning confocal microscope (Carl Zeiss AG). p-AKT(S473) (4060) and cleaved Caspase-3 (9664) antibodies were from Cell Signaling Technology. Rabbit polyclonal antibody against Ki67 (ab833) was from Abcam, and Mouse anti-laminin-5 (MAB1947) was from EMD Millipore. Alexa Fluor 488 anti-rabbit (A11008) and Alexa Fluor 594 anti-mouse (A21201) secondary antibodies, and ProLong® Gold antifade reagent with DAPI (P36931) were obtained from Life Technologies.
Pappas K., Xu J., Zairis S., Resnick-Silverman L., Abate F., Steinbach N., Ozturk S., Saal L.H., Su T., Cheung P., Schmidt H., Aaronson S., Hibshoosh H., Manfredi J., Rabadan R, & Parsons R. (2017). p53 maintains baseline expression of multiple tumor suppressor genes. Molecular cancer research : MCR, 15(8), 1051-1062.
+ Open protocol
+ Expand
2

Western Blot Analysis of Angiogenic Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysates were separated by SDS‐PAGE and electrophoretic transferred onto the PVDF (polyvinylidene difluoride) membranes (BIO‐RAD). Transferred membranes were blocked with 5% skim milk in TBS with 0.1% Tween‐20, and then incubated with primary antibodies at 4°C overnight. Following day, membranes were washed three times in TBS buffer with 0.1% Tween‐20 and incubated with the secondary antibody for 45 min at room temperature. Membranes were washed five times in TBS buffer with 0.1% Tween‐20 each 5 min then imaging by ChemiDoc Imaging system (BIO‐RAD). The following antibodies were purchased from Cell Signaling Technology [(VEGFR2‐ #9698, p‐AKT S473‐ #4060, p‐p44/42‐ #9101, p‐p38‐ #3871, p‐PLCγ2‐#4511, neuropilin 1‐ #3725)]. Antibodies to βIV‐spectrin and β‐actin were purchased from Santa Cruz Biotechnology (#SC514744) and Sigma‐Aldrich (#A1978) respectively. VEGFR1 antibody was purchased from R&D Systems (#AF471).
Kwak E., Ahmed T., Flores P.C., Ortiz H.R., Langlais P.R., Mythreye K, & Lee N.Y. (2023). Beta IV spectrin inhibits the metastatic growth of melanoma by suppressing VEGFR2‐driven tumor angiogenesis. Cancer Medicine, 12(18), 18981-18987.
+ Open protocol
+ Expand
3

Immunohistochemical Analysis of Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tissues were fixed overnight in 4% paraformaldehyde, embedded in paraffin, and cut into 5 µm thick sections. Sections were subject to hematoxylin and eosin staining, and immunohistochemical staining following standard protocols. The following primary antibodies were used: pERKT202/Y204 (4370) and pAKTS473 (4060) (Cell signaling), and pFRS2Y436 (ab193363) (Abcam).
Manchado E., Weissmueller S., Morris JP I.V., Chen C.C., Wullenkord R., Lujambio A., de Stanchina E., Poirier J.T., Gainor J.F., Corcoran R.B., Engelman J.A., Rudin C.M., Rosen N, & Lowe S.W. (2016). A combinatorial strategy for treating KRAS mutant lung cancer. Nature, 534(7609), 647-651.
+ Open protocol
+ Expand
4

Western Blot Analysis of Angiogenic Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysates were separated by SDS‐PAGE and electrophoretic transferred onto the PVDF (polyvinylidene difluoride) membranes (BIO‐RAD). Transferred membranes were blocked with 5% skim milk in TBS with 0.1% Tween‐20, and then incubated with primary antibodies at 4°C overnight. Following day, membranes were washed three times in TBS buffer with 0.1% Tween‐20 and incubated with the secondary antibody for 45 min at room temperature. Membranes were washed five times in TBS buffer with 0.1% Tween‐20 each 5 min then imaging by ChemiDoc Imaging system (BIO‐RAD). The following antibodies were purchased from Cell Signaling Technology [(VEGFR2‐ #9698, p‐AKT S473‐ #4060, p‐p44/42‐ #9101, p‐p38‐ #3871, p‐PLCγ2‐#4511, neuropilin 1‐ #3725)]. Antibodies to βIV‐spectrin and β‐actin were purchased from Santa Cruz Biotechnology (#SC514744) and Sigma‐Aldrich (#A1978) respectively. VEGFR1 antibody was purchased from R&D Systems (#AF471).
Kwak E., Ahmed T., Flores P.C., Ortiz H.R., Langlais P.R., Mythreye K, & Lee N.Y. (2023). Beta IV spectrin inhibits the metastatic growth of melanoma by suppressing VEGFR2‐driven tumor angiogenesis. Cancer Medicine, 12(18), 18981-18987.
+ Open protocol
+ Expand
5

Immunohistochemical Analysis of Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tissues were fixed overnight in 4% paraformaldehyde, embedded in paraffin, and cut into 5 µm thick sections. Sections were subject to hematoxylin and eosin staining, and immunohistochemical staining following standard protocols. The following primary antibodies were used: pERKT202/Y204 (4370) and pAKTS473 (4060) (Cell signaling), and pFRS2Y436 (ab193363) (Abcam).
Manchado E., Weissmueller S., Morris JP I.V., Chen C.C., Wullenkord R., Lujambio A., de Stanchina E., Poirier J.T., Gainor J.F., Corcoran R.B., Engelman J.A., Rudin C.M., Rosen N, & Lowe S.W. (2016). A combinatorial strategy for treating KRAS mutant lung cancer. Nature, 534(7609), 647-651.
+ Open protocol
+ Expand
6

Rictor CRISPR/Cas9 KO and RRM1 Overexpression

Check if the same lab product or an alternative is used in the 5 most similar protocols
Rictor CRISPR/Cas9 KO plasmid (sc-400710-KO-2) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rictor shRNAs (#1853 and #1854) were obtained from Addgene (Cambridge, MA). Flag-RRM1 plasmid was purchased from Genscript (Nanjing, China). Anti-RRM1(sc-377415), anti-RRM2(sc-398294) anti-β-Actin(sc-47778) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). p-Akt substrate (RXXS/T) (#9614), BrdU (#5292), Ki67(#9027), Cleaved caspase-3 (#9579) and pAkt S473 (#4060) antibodies were purchased from Cell Signaling Technology (Danvers, MA). γH2AX (05-636) was purchased from Millipore (Billerica, MA). Rictor (A300-459A), pRPA2 S33 (A300-246A), RPA2 (A300-244A) were purchased from Bethyl Laboratories (Montgomery, TX). Gemcitabine, PP242 and Rapamycin were obtained from Selleckchem (Houston, TX).
Tian L., Chen C., Guo Y., Zhang F., Mi J., Feng Q., Lin S., Xi N., Tian J., Yu L., Chen Y., Cao M., Lai C., Fan J., Zhang Y, & Chen G. (2021). mTORC2 regulates ribonucleotide reductase to promote DNA replication and gemcitabine resistance in non-small cell lung cancer. Neoplasia (New York, N.Y.), 23(7), 643-652.
+ 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!