Calcineurin cellular assay kit

Manufactured by Enzo Life Sciences

Sourced in United States

The Calcineurin cellular assay kit is a laboratory tool designed to measure the activity of the calcineurin enzyme in cellular samples. Calcineurin is a key regulator of various cellular processes, and this kit provides a standardized method for quantifying its enzymatic activity.

Automatically generated - may contain errors

Lab products found in correlation

Adenosine diphosphate (adp), by Helena Laboratories (1 mentions) Alexa 488 conjugated fibrinogen, by Thermo Fisher Scientific (1 mentions) Pe conjugated jon a antibody, by Emfret Analytics (1 mentions)

Close spelling variants of this product

Calcineurin cellular activity assay kit Calcineurin

2 protocols using calcineurin cellular assay kit

Quantification of Cellular Calcineurin Activity

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

Cellular calcineurin activity was measured by calcineurin cellular assay kit (ENZO Life Sciences, Plymouth Meeting, PA, USA) according to the manufacturer's protocol. Briefly, treated cells were collected using the lysis buffer supplied in the kit. The extracts were then subjected to desalting resin to remove free phosphate. The desalted lysates were incubated with the calcineurin substrate RII phosphopeptide in assay buffer and EGTA buffer at 30°C for 30 min to determine total phosphatase activity and phosphatase activity without calcineurin, respectively. Then, 100 μl BIOMOL GREEN was added to each well to visualise the phosphate release. The colour was allowed to develop at room temperature for 20 min before the detection of the absorbance value at 620 nm on the microplate spectrophotometer. The absorbance value A₆₂₀, _total –A_{620, EGTA buffer} was converted to phosphate release using the standard curve that was prepared in each assay. The phosphate release was divided by protein concentration and normalised to the control group to obtain relative calcineurin activity.

He H., Liu X., Lv L., Liang H., Leng B., Zhao D., Zhang Y., Du Z., Chen X., Li S., Lu Y, & Shan H. (2014). Calcineurin suppresses AMPK-dependent cytoprotective autophagy in cardiomyocytes under oxidative stress. Cell Death & Disease, 5(1), e997-.

+ Open protocol

+ Expand

Platelet Activation Regulation by Calcineurin

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

ADP and collagen were purchased from Helena Laboratories (Beaumont, TX), while thrombin was from Hematologic Technologies Inc. (Essex Junction, VT). Protease activated receptor 4 –activating peptide (PAR4-AP) AYPGKF was synthesized by the Protein Core at Baylor College of Medicine (BCM). CsA and FK506 were from Calbiochem-EMD Millipore (Darmstadt, Germany). Alexa 488-conjugated fibrinogen was from Invitrogen-life Technologies (Grand Island, NY). Human fibrinogen was from Enzyme Research Laboratories Inc. (South bend, IN). Fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD62 (P-selectin), Phycoerythrin (PE)–conjugated anti-mouse CD41 (αIIb) and control IgG tagged to PE or FITC were purchased from BD Bioscience (San Jose, CA). PE-conjugated JON/A antibody was from Emfret Analytics (Eibelstadt, Germany). Antibodies to phospho Src Tyr⁴¹⁸, Src, phospho p38 (Thr¹⁸⁰ Tyr¹⁸²), p38, phospho filamin (Ser²¹⁵²), and filamin were from Cell Signaling Technologies (Beverly, MA). Antibodies to PP2B-Aα, PP2B-Aβ were from Santa Cruz Biotechnology (Santa Cruz, CA). Calcineurin cellular assay kit was from Enzo Life Sciences, Inc. (Farmingdale, NY). A preformed mix of four independent (SMART pool) siRNAs targeting human PP2B-Aβ and non-specific control siRNA pool were purchased from Dharmacon (Thermo Fisher Scientific, Lafayette, CO).

Khatlani T., Pradhan S., Da Q., Gushiken F.C., Bergeron A.L., Langlois K.W., Molkentin J.D., Rumbaut R.E, & Vijayan K.V. (2014). The β isoform of the catalytic subunit of protein phosphatase 2B restrains platelet function by suppressing outside-in αIIbβ3 integrin signaling. Journal of thrombosis and haemostasis : JTH, 12(12), 2089-2101.

+ 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!