Certificate of Analysis

ProQinase™ MAPKAPK5 (non activated) mitogen-activated -activated protein kinase 5 RecombinantProteins

Recombinant Protein Kinase MAPKAPK5 Lot 001: Western blot analysis MAPKAPK5 HGNC Symbol: anti-MAPKAPK5 anti-GST Synonyms: PRAK kDa kDa 250 250 150 150 Product No.: 0380-0000-1 100 100 GST- GST- 75 75 Lot: 001 MAPKAPK5 MAPKAPK5

50 50 Description: Human MAPKAPK5 Amino acids M1-Q473 (as in NCBI/Protein entry 37 37 NP_620777.1)*, N-terminally fused to GST- 25 25 HIS6-Thrombin cleavage site 20 20 15 15 Product identity: MAPKAPK5, Lot 001, was confirmed as MAPKAPK5 by specific Western 500 ng GST-MAPKAPK5 500 ng GST-MAPKAPK5 Blotting using anti MAPKAPK5 antibody Kinase activity MAPKAPK5 (not activated) vs

Theoretical MWFusion Protein: 84,116 Da active MAPKAPK5:

20000 Expression: Baculovirus infected Sf9 cells n = 3 Substrate (µg/well): : + 1: no substrate : - Enzyme 2: 1 Purification: One-step affinity purification using 15000 3: 2 4: 4 GSH-agarose

10000 Storage buffer: 50 mM Tris-HCl, pH 8.0;

100 mM NaCl, 5 mM DTT, 4 mM reduced 5000

glutathione, 20% glycerol (cpm) activity Kinase

0 1 2 3 4 1 2 3 4 1 2 3 4 Storage temperature: -80°C 25 50 100 Avoid repeated freeze-thaw cycles! Kinase (ng/well) Protein concentration: 0.113 µg/µl (Bradford method using BSA [Sigma, cat# 20000 Substrate (µg/well): : + Enzyme n = 3 A-7638, Lot 79H7641] as standard protein) 1: no substrate : - Enzyme 2: 1 15000 3: 2 4: 4

Coomassie stain: 10000

kDa 5000

250 (cpm) activity Kinase

150 0 1 2 3 4 1 2 3 4 1 2 3 4 25 50 100 100 GST- 75 MAPKAPK5 Kinase (ng/well) Final assay concentrations: - 60 mM HEPES-NaOH, pH 7.5 50 - 3 mM MgCl2

37 - 3 mM MnCl2 - 3 µM Na-orthovanadate - 1.2 mM DTT 25 - 50 µg / ml PEG 20 20.000 - 1 µM ATP (798,000 cpm 33P-γ-ATP) 15 - Substrate (variable): ATF2 - Recombinant MAPKAPK5 (not activated) or active 2.0 µg GST-MAPKAPK5 MAPKAPK5 (variable) Assay: 33PanQinase® Assay

This product was manufactured at Reaction Biology in Freiburg, Germany, and is for in vitro research use only, not for use in or animals. © European Union, 2020. Material may not be reproduced or distributed without written permission from Reaction Biology Europe GmbH

Reaction Biology www.reactionbiology.com [email protected] Certificate of Analysis - Page 2 -

ProQinase™ MAPKAPK5 Recombinant Product No.: 0380-0000-1

MAPKAPK5 Recombinant Fusion Protein Amino Acid Sequence 1 MSPILGYWKI KGLVQPTRLL LEYLEEKYEE HLYERDEGDK WRNKKFELGL EFPNLPYYID 60 61 GDVKLTQSMA IIRYIADKHN MLGGCPKERA EISMLEGAVL DIRYGVSRIA YSKDFETLKV 120 121 DFLSKLPEML KMFEDRLCHK TYLNGDHVTH PDFMLYDALD VVLYMDPMCL DAFPKLVCFK 180 181 KRIEAIPQID KYLKSSKYIA WPLQGWQATF GGGDHPPKSD PMGHHHHHHG RRRASVAAGI 240 241 LVPRGSPGLD GIYARGIQAS MSEESDMDKA IKETSILEEY SINWTQKLGA GISGPVRVCV 300 301 KKSTQERFAL KILLDRPKAR NEVRLHMMCA THPNIVQIIE VFANSVQFPH ESSPRARLLI 360 361 VMEMMEGGEL FHRISQHRHF TEKQASQVTK QIALALRHCH LLNIAHRDLK PENLLFKDNS 420 421 LDAPVKLCDF GFAKIDQGDL MTPQFTPYYV APQVLEAQRR HQKEKSGIIP TSPTPYTYNK 480 481 SCDLWSLGVI IYVMLCGYPP FYSKHHSRTI PKDMRRKIMT GSFEFPEEEW SQISEMAKDV 540 541 VRKLLKVKPE ERLTIEGVLD HPWLNSTEAL DNVLPSAQLM MDKAVVAGIQ QAHAEQLANM 600 601 RIQDLKVSLK PLHSVNNPIL RKRKLLGTKP KDSVYIHDHE NGAEDSNVAL EKLRDVIAQC 660 661 ILPQAGKGEN EDEKLNEVMQ EAWKYNRECK LLRDTLQSFS WNGRGFTDKV DRLKLAEIVK 720 721 QVIEEQTTSH ESQ 780 1-218: GST Red: HIS6-tag Pink: Thrombin blue: MAPKAPK5

MAPKAPK5 wt1 amino acid sequence 1 MSEESDMDKA IKETSILEEY SINWTQKLGA GISGPVRVCV KKSTQERFAL KILLDRPKAR 60 61 NEVRLHMMCA THPNIVQIIE VFANSVQFPH ESSPRARLLI VMEMMEGGEL FHRISQHRHF 120 121 TEKQASQVTK QIALALRHCH LLNIAHRDLK PENLLFKDNS LDAPVKLCDF GFAKIDQGDL 180 181 MTPQFTPYYV APQVLEAQRR HQKEKSGIIP TSPTPYTYNK SCDLWSLGVI IYVMLCGYPP 240 241 FYSKHHSRTI PKDMRRKIMT GSFEFPEEEW SQISEMAKDV VRKLLKVKPE ERLTIEGVLD 300 301 HPWLNSTEAL DNVLPSAQLM MDKAVVAGIQ QAHAEQLANM RIQDLKVSLK PLHSVNNPIL 360 361 RKRKLLGTKP KDSVYIHDHE NGAEDSNVAL EKLRDVIAQC ILPQAGKGEN EDEKLNEVMQ 420 421 EAWKYNRECK LLRDTLQSFS WNGRGFTDKV DRLKLAEIVK QVIEEQTTSH ESQ 480 blue: MAPKAPK5 sequence expressed in fusionprotein

1NCBI/Protein accession number NP_620777.1

This product was manufactured at Reaction Biology in Freiburg, Germany, and is for in vitro research use only, not for use in humans or animals. © European Union, 2020. Material may not be reproduced or distributed without written permission from Reaction Biology Europe GmbH

Reaction Biology www.reactionbiology.com [email protected]