Name: Recombinant Human Active ZAP70 Protein, CF
Brand: Novus Biologicals
SKU: 3709-KS
Price: 560 USD

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Publications for ZAP70 (3709-KS)(2)

We have publications tested in 1 confirmed species: Mouse.

We have publications tested in 1 application: Bioassay.

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Bioassay

(2)

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Showing Publications 1 - 2 of 2.

Publications using 3709-KS	Applications	Species
ML Giardino T, D Dutta, PR Mittelstad, J Guha, MM Gaida, K Fish, VA Barr, IO Akpan, LE Samelson, HD Tagad, S Debnath, LM Miller Jen, E Appella, JD Ashwell Intensity and duration of TCR signaling is limited by p38 phosphorylation of ZAP-70and destabilization of the signalosome Proc. Natl. Acad. Sci. U.S.A., 2018-02-12;0(0):. 2018-02-12 [PMID: 29440413] (Bioassay)	Bioassay
MS Alam, MM Gaida, S Debnath, HD Tagad, LM Miller Jen, E Appella, MJ Rahman, JD Ashwell Unique properties of TCR-activated p38 are necessary for NFAT-dependent T-cell activation PLoS Biol., 2018-01-22;16(1):e2004111. 2018-01-22 [PMID: 29357353] (Bioassay, Mouse)	Bioassay	Mouse

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Reactivity	HuSpecies Glossary
Applications	Bioactivity
Format	Carrier-Free

Details of Functionality	The specific activity of ZAP70 was determined to be 13 nmol/min/mg using a Poly (Glu:Tyr, 4:1) synthetic peptide substrate.
Source	Spodoptera frugiperda, Sf 9 (baculovirus)-derived human ZAP70 protein
Accession #	NM_001079
N-terminal Sequence	Using an N-terminal GST tag
Protein/Peptide Type	Recombinant Proteins
Gene	ZAP70
Purity	>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Storage	This product is stable at ≤ ‑70 °C for up to 1 year from the date of receipt. For optimal storage, aliquot into smaller quantities after centrifugation and store at recommended temperature. Avoid repeated freeze-thaw cycles.
Buffer	Supplied in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM Glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, 25% Glycerol.
Purity	>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Assay Procedure	Active Kinase - Active ZAP70 (0.1 μg/μL) diluted with Kinase Dilution Buffer X and assayed as outlined in sample activity plot. Note: These are suggested working dilutions and it is recommended that the researcher perform a serial dilution of Active ZAP70 for optimal results. Kinase Assay Buffer III (5X) - 200 mM Tris-HCl, pH 7.4, 100 mM MgCl₂, and 0.5 mg/mL BSA. Add fresh DTT prior to use to a final concentration of 250 μM. Kinase Dilution Buffer X - 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 2.5 mM MnCl₂, and 0.1 mg/mL BSA. Add fresh DTT prior to use to a final concentration of 50 μM. ADP-Glo™ Kinase Assay Kit - 10 mM of ATP Solution, 10 mM of ADP Solution, ADP-Glo™ Reagent, and Kinase Detection Reagent. Substrate - Poly (4:1 Glu:Tyr) synthetic peptide substrate diluted in 25 mM Tris-HCl to a final concentration of 1 mg/mL. Cofactor: 2.5 M MnCl₂ - Diluted in distilled water to a working concentration of 1 M. Thaw the Active ZAP70, Kinase Assay Buffer III (5X), and Substrate on ice. Prepare a 15 μL enzyme dilution with Kinase Dilution Buffer X at the desired concentration, in a pre-chilled 96-well plate. Prepare a Substrate/ATP mixture as follows (25 μM ATP example): a. 10 mM ATP Solution: 1 μL b. Kinase Assay Buffer III (5X): 78 μL c. Substrate at 1 mg/mL: 80 μL d. 1 M MnCl₂: 1 μL Transfer the following reaction components prepared in Steps 1 and 2 to a 384-well opaque plate, bringing the reaction volume up to 5 μL: a. Diluted Active ZAP70: 3 μL b. Substrate/ATP (mix as prepared in Step 2). This initiates the reaction: 2 μL Set up the blank control as outlined in Step 2, excluding the addition of the kinase. Replace the kinase with an equal volume of Kinase Dilution Buffer X. Incubate at ambient temperature for 40 minutes. After the 40 minute incubation period, terminate the reaction and deplete the remaining ATP by adding 5 μL of ADP-Glo™ Reagent. Spin down and shake the 384-well plate. Then incubate the reaction mixture for another 40 minutes at ambient temperature. Then add 10 μL of the Kinase Detection Reagent to the 384-well plate and incubate the reaction mixture for another 30 minutes at ambient temperature. Read the 384-well reaction plate using the Luminescence Module Protocol on a GloMax®-Multi Microplate Multimode Reader. Determine the corrected activity (RLU) by removing the blank control value (see step 4) for each sample and calculate the kinase specific activity as outlined below. Calculation of Specific Activity of ADP (RLU/pmol) From ADP standard curve, determine RLU/pmol of ADP Kinase Specific Activity (SA) (pmol/minutes/μg or nmol/minutes/mg) Corrected RLU from reaction / [(SA of ADP in RLU/pmol) x (Reaction time in minutes) x (Enzyme amount in μg or mg)]

Gene Symbol	ZAP70
Uniprot	NM_001079(Human)

Recombinant Human Active ZAP70 Protein, CF

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