PV6289 Certificate of Analysis for Lot 1868200

Certificate of Analysis CDK2/cyclin A1, 10 µg Recombinant human full-length protein

5791 Van Allen Way Part Number: PV6289 Carlsbad, CA 92008 Lot Number: 1868200 Phone: 760.603.7200 Immediate Storage: -80°C Fax: 760.602.6500 Shipping Conditions: dry ice www.lifetechnologies.com

Description: Storage and Handling: Recombinant human full-length active CDK2/cyclin A1 were co-expressed For maximum recovery please spin prior to use. Unless noted below, in insect cells using N−terminal GST tags. aliquots of the 5 ug, 10ug and 20ug sizes of kinase are not recommended CDK2 is a member of the Cyclin-Dependent Kinase family that is as materials can be used in original packaging until exhausted. For larger ubiquitously expressed. CDK2 is a catalytic subunit of the cyclin-dependent sizes, the number of freeze/thaws may be reduced by preparing aliquots, protein kinase complex, whose activity is restricted to the G1-S phase, and aliquots below 20 μL are not recommended. Please never store a kinase essential for cell cycle G1/S phase transition. CDK2 associates with and is diluted. If properly stored at −80°C, this product is guaranteed for 6 months regulated by the regulatory subunits of the complex including Cyclin A or E, from date of purchase. CDK inhibitor p21Cip1 (CDKN1A) and p27Kip1 (CDKN1B). CDK2 Storage Buffer: phosphorylates multiple cellular substrates including SMAD3 and FOXO1. 50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 10 mM glutathione, Phosphorylation of FOXO1 leads to its inhibition. 0.1 mM EDTA, 0.25 mM DTT, 0.1mM PMSF and 25% Glycerol. Accession Numbers: The gene accession number for the subunits (CDK2/Cyclin A1) are NP_001789.2 and NP_001104515.1.. Specific Activity: 74 nmoles of phosphate transferred to Histone H1 per minute per mg of total protein at 30°C. Concentration: 0.1 mg/mL total protein as measured using the Bradford protein assay with BSA as a standard. Calculated 721 nM. Aliases: CDK2: p33(CDK2) CyclinA1: CCNA1

QUALITY ASSURANCE

CDK2/cyclin A1 Activity Graph Gel Information for CDK2/cyclin A1

Page Description: Run on an SDS-PAGE gel and stained with Coomassie®.

Lane 1: Molecular Weight markers as labeled.

Lane 2: CDK2/cyclin A1

Dilution Buffer:

5 mM MOPS (pH 7.2), 2.5 mM β−glycerol−phosphate, 5 mM MgCl2, Purity: 1 mM EGTA, 0.4 mM EDTA and 50 ng/μL BSA. > 80% as determined by a Coomassie® blue stained SDS-PAGE gel. Assay Conditions: Molecular Weight: CDK2/cyclin A1 was pre-diluted in enzyme dilution buffer and assayed in 60.2 / 78.5 kDa. Calculated from the protein sequence(s). 5 mM MOPS (pH 7.2), 2.5 mM β−glycerol-phosphate, 5 mM MgCl2, 0.4 mM EDTA, 1 mM EGTA, 0.05 mM DTT, with 50 μM ATP, trace [33P]-γ- ATP and 200 μg/mL Histone H1 for 15 minutes at 30°C.

For questions, please contact our Technical Support Team N. Am Ph#: 800-955-6288 or INTL Ph#: 760-603-7200 Select option 5, ext. 40266 Email: [email protected] Printed 08/Feb/2017 Version 1.0 Page 1 Certificate of Analysis Part Number: PV6289, Lot Number: 1868200

Protein sequence alignment with reference sequence(s)

GenBank Accession Number: NP_001789.2

CDK2

1 MSPILGYWKI KGLVQPTRLL LEYLEEKYEE HLYERDEGDK WRNKKFELGL EFPNLPYYID GDVKLTQSMA IIRYIADKHN MLGGCPKERA EISMLEGAVL GST TAG 1 MSPILGYWKI KGLVQPTRLL LEYLEEKYEE HLYERDEGDK WRNKKFELGL EFPNLPYYID GDVKLTQSMA IIRYIADKHN MLGGCPKERA EISMLEGAVL IVGN CDK2 1 ------NP_001789.2

101 DIRYGVSRIA YSKDFETLKV DFLSKLPEML KMFEDRLCHK TYLNGDHVTH PDFMLYDALD VVLYMDPMCL DAFPKLVCFK KRIEAIPQID KYLKSSKYIA 101 DIRYGVSRIA YSKDFETLKV DFLSKLPEML KMFEDRLCHK TYLNGDHVTH PDFMLYDALD VVLYMDPMCL DAFPKLVCFK KRIEAIPQID KYLKSSKYIA 1 ------

201 WPLQGWQATF GGGDHPPKSD LVPRGS 201 WPLQGWQATF GGGDHPPKSD LVPRGSMENF QKVEKIGEGT YGVVYKARNK LTGEVVALKK IRLDTETEGV PSTAIREISL LKELNHPNIV KLLDVIHTEN 1 ------MENF QKVEKIGEGT YGVVYKARNK LTGEVVALKK IRLDTETEGV PSTAIREISL LKELNHPNIV KLLDVIHTEN

226 301 KLYLVFEFLH QDLKKFMDAS ALTGIPLPLI KSYLFQLLQG LAFCHSHRVL HRDLKPQNLL INTEGAIKLA DFGLARAFGV PVRTYTHEVV TLWYRAPEIL 75 KLYLVFEFLH QDLKKFMDAS ALTGIPLPLI KSYLFQLLQG LAFCHSHRVL HRDLKPQNLL INTEGAIKLA DFGLARAFGV PVRTYTHEVV TLWYRAPEIL

226 401 LGCKYYSTAV DIWSLGCIFA EMVTRRALFP GDSEIDQLFR IFRTLGTPDE VVWPGVTSMP DYKPSFPKWA RQDFSKVVPP LDEDGRSLLS QMLHYDPNKR 175 LGCKYYSTAV DIWSLGCIFA EMVTRRALFP GDSEIDQLFR IFRTLGTPDE VVWPGVTSMP DYKPSFPKWA RQDFSKVVPP LDEDGRSLLS QMLHYDPNKR

226 501 ISAKAALAHP FFQDVTKPVP HLRL 275 ISAKAALAHP FFQDVTKPVP HLRL

GenBank Accession Number: NP_001104515.1

Cyclin A1

201 WPLQGWQATF GGGDHPPKSD LVPRGS 201 WPLQGWQATF GGGDHPPKSD LVPRGSMETG FPAIMYPGSF IGGWGEEYLS WEGPGLPDFV FQQPVESEAM HCSNPKSGVV LATVARGPDA CQILTRAPLG 1 ------METG FPAIMYPGSF IGGWGEEYLS WEGPGLPDFV FQQPVESEAM HCSNPKSGVV LATVARGPDA CQILTRAPLG

226 301 QDPPQRTVLG LLTANGQYRR TCGQGITRIR CYSGSENAFP PAGKKALPDC GVQEPPKQGF DIYMDELEQG DRDSCSVREG MAFEDVYEVD TGTLKSDLHF 75 QDPPQRTVLG LLTANGQYRR TCGQGITRIR CYSGSENAFP PAGKKALPDC GVQEPPKQGF DIYMDELEQG DRDSCSVREG MAFEDVYEVD TGTLKSDLHF

226 401 LLDFNTVSPM LVDSSLLSQS EDISSLGTDV INVTEYAEEI YQYLREAEIR HRPKAHYMKK QPDITEGMRT ILVDWLVEVG EEYKLRAETL YLAVNFLDRF 175 LLDFNTVSPM LVDSSLLSQS EDISSLGTDV INVTEYAEEI YQYLREAEIR HRPKAHYMKK QPDITEGMRT ILVDWLVEVG EEYKLRAETL YLAVNFLDRF

226 501 LSCMSVLRGK LQLVGTAAML LASKYEEIYP PEVDEFVYIT DDTYTKRQLL KMEHLLLKVL AFDLTVPTTN QFLLQYLRRQ GVCVRTENLA KYVAELSLLE 275 LSCMSVLRGK LQLVGTAAML LASKYEEIYP PEVDEFVYIT DDTYTKRQLL KMEHLLLKVL AFDLTVPTTN QFLLQYLRRQ GVCVRTENLA KYVAELSLLE

226 601 ADPFLKYLPS LIAAAAFCLA NYTVNKHFWP ETLAAFTGYS LSEIVPCLSE LHKAYLDIPH RPQQAIREKY KASKYLCVSL MEPPAVLLLQ 375 ADPFLKYLPS LIAAAAFCLA NYTVNKHFWP ETLAAFTGYS LSEIVPCLSE

* highlighted residues denote differences from the reference protein sequence(s).

Tony Goossens, Engineer II, QA/QC Date: 07/Oct/2014

Coomassie® is a registered trademark of Imperial Chemical Industries.

For Research Use Only. Not for use in diagnostic procedures.

For questions, please contact our Technical Support Team N. Am Ph#: 800-955-6288 or INTL Ph#: 760-603-7200 Select option 5, ext. 40266 Email: [email protected] Printed 08/Feb/2017 Version 1.0 Page 2