Cannot open database "Origene_clones" requested by the login. The login failed.
Login failed for user 'OrigeneClones'.System.Data.SqlClient.SqlException: Cannot open database "Origene_clones" requested by the login. The login failed.
Login failed for user 'OrigeneClones'.
at System.Data.SqlClient.SqlInternalConnection.OnError(SqlException exception, Boolean breakConnection)
at System.Data.SqlClient.TdsParser.ThrowExceptionAndWarning(TdsParserStateObject stateObj)
at System.Data.SqlClient.TdsParser.Run(RunBehavior runBehavior, SqlCommand cmdHandler, SqlDataReader dataStream, BulkCopySimpleResultSet bulkCopyHandler, TdsParserStateObject stateObj)
at System.Data.SqlClient.SqlInternalConnectionTds.CompleteLogin(Boolean enlistOK)
at System.Data.SqlClient.SqlInternalConnectionTds.AttemptOneLogin(ServerInfo serverInfo, String newPassword, Boolean ignoreSniOpenTimeout, Int64 timerExpire, SqlConnection owningObject)
at System.Data.SqlClient.SqlInternalConnectionTds.LoginNoFailover(String host, String newPassword, Boolean redirectedUserInstance, SqlConnection owningObject, SqlConnectionString connectionOptions, Int64 timerStart)
at System.Data.SqlClient.SqlInternalConnectionTds.OpenLoginEnlist(SqlConnection owningObject, SqlConnectionString connectionOptions, String newPassword, Boolean redirectedUserInstance)
at System.Data.SqlClient.SqlInternalConnectionTds..ctor(DbConnectionPoolIdentity identity, SqlConnectionString connectionOptions, Object providerInfo, String newPassword, SqlConnection owningObject, Boolean redirectedUserInstance)
at System.Data.SqlClient.SqlConnectionFactory.CreateConnection(DbConnectionOptions options, Object poolGroupProviderInfo, DbConnectionPool pool, DbConnection owningConnection)
at System.Data.ProviderBase.DbConnectionFactory.CreatePooledConnection(DbConnection owningConnection, DbConnectionPool pool, DbConnectionOptions options)
at System.Data.ProviderBase.DbConnectionPool.CreateObject(DbConnection owningObject)
at System.Data.ProviderBase.DbConnectionPool.UserCreateRequest(DbConnection owningObject)
at System.Data.ProviderBase.DbConnectionPool.GetConnection(DbConnection owningObject)
at System.Data.ProviderBase.DbConnectionFactory.GetConnection(DbConnection owningConnection)
at System.Data.ProviderBase.DbConnectionClosed.OpenConnection(DbConnection outerConnection, DbConnectionFactory connectionFactory)
at linked_ascx_related_5_button_new.GetProteinBtn(HyperLink lnkPotein)Cannot open database "Origene_clones" requested by the login. The login failed.
Login failed for user 'OrigeneClones'.
1 vial of 100 µg gene specific transient over-expression cell lysate in RIPA buffer
1 vial of 100 µg empty vector transfected control cell lysate in RIPA buffer
1 vial of 250ul 2xSDS Sample Buffer (4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromphenol blue, 100mM DTT)
The lysate is shipped with dry ice. Upon receiving, store the sample at -20ºC. Lysate samples are stable for 12 months from date of receipt when stored at -20ºC. Avoid repeated freeze-thaw cycles. Lysate samples can be diluted with 2xSDS Sample Buffer provided. After dilution, the protein sample should be aliquoted and stored at -20ºC for long term storage. Prior to SDS-PAGE fractionation, boil the lysate for 5 minutes.
HEK293T cells in 10-cm dishes were transiently transfected with MegaTran Transfection Reagent (TT200002) and 5ug TrueORF cDNA plasmid. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix (Sigma), 1mM PMSF and 1mM Na3VO4), and then centrifuged to clarify the lysate. Protein concentration was measured by BCA kit (Thermo Scientific Inc.). Cell lysates were aliquoted and stored at -20ºC before shipping.
Western validation with an anti-DDK antibody* L: Control HEK293 lysate R: Over-expression lysate
*: DDK-tag is the same as FLAG tag. Flag® is a registered trademark of Sigma-Aldrich
**: Delivery time is an estimate in business days. Occasional delays may occur due to unforeseen complexities in the preparation of your construct. International customers may expect an additional 1-2 weeks in shipping