Histidine (His) Tag Plasmids Vectors Information

His tag properties and uses

 

Histidine tags (His tags) can be easily encoded within DNA sequences and be positioned at either end of the protein. They can be used both for purification of proteins and also for detection by western blotting. Traditionally purification has involved adsorption of the protein to nickel or cobalt chromatography columns, followed by elution with a base. However anti-His antibodies are efficient for detection and can also be used successfully for purification in a range of formats.

 

Most his tags are hexahistidine (His6) although His10 is also available, and binds more strongly to the nickel and cobalt columns. His tags are suitable for purification of many proteins, including those that need to be purified denatured.

 

Histidine has a weak positive charge at neutral pH. His6 has a molecular weight of 930 Da, while His10 is 1550 Da.

 

Purification of proteins using hexahistidine: the principle

A typical hexahistidine purification is as follows:

  1. Harvest and lyse host cells with expressed histidine-tagged protein, and clarify by centrifugation.
  2. Incubate the lysate with a slurry of Nickel beads (eg Ni-Sepharose). I ml of 50% slurry can bind about 20 mg of His-tagged protein.
  3. Pack the slurry into an empty disposable PD10 column.

His

Our product range

Oxford Genetics provides a broad range of plasmids containing N-terminal and C-terminal His tags, both His6 and His10. In some plasmids the tags flank reporter genes, while in others they are positioned adjacent the MCS enabling you to insert your own gene in frame as required.

 

The plasmid structure shown contains a hexahistidine tag positioned downstream of the MCS, allowing simple inclusion of the tag at the C terminus of your gene of interest, driven by the CMV promoter. The structure also includes an Enterokinase cleavage site for simple removal of the tag following protein production, if required. The plasmid also contains puromycin resistance, driven by the ubiquitin promoter, for selection in mammalian cells, together with kanamycin resistance for selection in bacteria. Our online catalogue contains a diverse array of plasmid structures, accessible via the buttons below.

 

Our Plasmid Builder facility also provides a simple means for you to design an efficient strategy for any modifications or cloning manipulations to these plasmids. Finally, as always, although we have designed these plasmids for simple and efficient cloning, we are happy to undertake any cloning steps that you prefer to outsource. You can access all these options through the Plasmid Builder button below.

 

Plasmids Introducing Functional Tags onto Your Gene of Interest (for use in mammalian cells)

N Terminal Tag Plasmids are in Blue

C Terminal Tag Plasmids are in Red

 

Protease Cleavage Site

For removal of tag after purification or analysis of your protein

none

EKT

TEV

FXa

Thrombin

3C

 

Epitope Tags

V5

OG3212 OG3422

OG3202

OG3411

OG3194

OG3404

OG3191

OG3400

OG3192

OG3401

OG3196

OG3405

Ha

OG3215

OG3425

OG3200

OG3409

OG1127

OG1128

OG1067

OG1068

OG1087

OG1088

OG1145

OG1146

C-Myc

OG3214

OG3424

OG3199

OG3408

OG1126

OG1125

OG1065

OG1066

OG1085

OG1086

OG1143

OG1144

S-Tag

OG3219

OG3429

OG1163

OG1164

OG1135

OG1136

OG1075

OG1076

OG1095

OG1096

OG1153

OG1154

T7

OG3218

OG3428

OG1161

OG1162

OG1133

OG1134

OG1073

OG1074

OG1093

OG1094

OG1151

OG1152

FLAG

OG3213

OG3423

OG3198

OG3407

OG1123

OG1124

OG1063

OG1064

OG1083

OG1084

OG1141

OG1142

Affinity Tags

MBP

OG3220

OG3430

OG1165

OG1166

OG3195

OG3403

OG1077

OG1078

OG1097

OG1098

OG1155

OG1156

GST

OG3216

OG3426

OG3201

OG3410

OG1129

OG1130

OG1069

OG1070

OG1089

OG1090

OG1147

OG1148

Strep

OG3217

OG3427

OG1159

OG1160

OG1131

OG1132

OG1071

OG1072

OG1091

OG1092

OG1149

OG1150

6His

OG3211

OG3421

OG3197

OG3406

OG1121

OG1122

OG1061

OG1062

OG1081

OG1082

OG1139

OG1140

10His

OG3210

OG3420

OG1157

OG1158

OG1119

OG1120

OG1059

OG1060

OG1079

OG1080

OG1137

OG1138

Dual

Tags

 

V5 & 6His

OG3203

OG3209

OG3412

OG3419

OG3207

OG3417

OG3204

OG3413

OG3205

OG3415

OG3208

OG3418

 

HA & 6His

OG1179

OG1267

OG1180

OG1268

OG1235

OG1236

OG1187

OG1188

OG1203

OG1204

OG1251

OG1252

 

c-Myc & 6His

OG1265

OG1177

OG1266

OG1178

OG1233

OG1234

OG1185

OG1186

OG1201

OG1202

OG1249

OG1250

 

S-Tag & 6His

OG1171

OG1275

OG1172

OG1276

OG1243

OG1244

OG1195

OG1196

OG1211

OG1212

OG1259

OG1260

 

T7 & 6His

 

OG1169

OG1273

OG1170

OG1274

OG1241

OG1242

OG1193

OG1194

OG1209

OG1210

OG1257

OG1258

 

FLAG & 6His

OG1175

OG1263

OG1176

OG1264

OG1231

OG1232

OG1183

OG1184

OG1199

OG1200

OG1247

OG1248

 

MBP & 6His

OG1173

OG1277

OG1174

OG1278

OG1245

OG1246

OG1197

OG3414

OG1213

OG1214

OG1261

OG1262

 

GST & 6His

OG1181

OG1269

OG1182

OG1270

OG1237

OG1238

OG1189

OG1190

OG1205

OG1206

OG1253

OG1254

 

Strep & 6His

OG1167

OG1271

OG1168

OG1272

OG1239

OG1240

OG1191

OG1192

OG1207

OG1208

OG1255

OG1256

 

 

V5 Tags Plasmid Vector Information

V5 tag properties and uses

 

The V5 epitope tag is derived from a peptide present at the C terminus of the P and V proteins of simian virus 5. It contains 14 amino acids, from N to C: GKPIPNPLLGLDST.

 

Oxford Genetics provides a range of plasmid vectors containing V5 tags in a range of orientations, to meet all your cloning requirements. Please browse through our products using the ‘Browse Plasmids’ menu. For example we provide V5 at both N and C sides of the MCS, allowing versatile positioning of your gene, we sell it with and without enzyme cleavage sites (for simple removal of the tag, if required, after protein production), and we sell it in a range of configurations with affinity tags (such as hexahistidine) to give you maximum flexibility.

 

The V5 tag is an epitope tag, meaning there are good antibodies that recognise it specifically. This makes it especially useful for immunoprecipitation, immunofluorescence and western blotting.

 

Immunoprecipitation of proteins using V5 tags: the principle

A typical V5 immunoprecipitation is as follows:

  1. Harvest and lyse host cells with expressed V5-tagged protein, and clarify by centrifugation

  2. Incubate the lysate with a slurry of gel beads bearing V5-capture antibodies, centrifuge and wash carefully.

  3. Elute the bound V5-tagged protein and its binding partner(s) with a buffer containing the free V5-elution peptide GKPIPNPLLGLDST.

 

Our product range

Oxford Genetics provides a broad range of plasmids containing N-terminal and C-terminal V5 tags. In some plasmids the tags flank reporter genes, while in others they are positioned adjacent the MCS enabling you to insert your own gene in frame as required.

 

The plasmid structure shown contains the V5 tag positioned downstream of the MCS, for inclusion at the C terminal of your inserted gene. The plasmid also contains puromycin resistance, driven by the ubiquitin promoter, for selection in mammalian cells, and kanamycin resistance for selection in bacteria.

 

Our Plasmid Builder facility also provides a simple means for you to design an efficient strategy for any modifications or cloning manipulations to these plasmids. Finally, as always, although we have designed these plasmids for simple and efficient cloning, we are happy to undertake any cloning steps that you prefer to outsource. You can access all these options through the Plasmid Builder button below.

 

 

Thioredoxin tag properties and uses

Thioredoxin tag properties and uses

 

The Thioredoxin (TRX) tag is used predominantly to increase the solubility and thermal stability of proteins expressed in bacterial systems, where it also assists in the refolding of proteins requiring a reducing environment. It also provides a useful epitope tag, and has a molecular weight of approximately 12 kDa.

 

Oxford Genetics provides a range of plasmid vectors containing TRX tags for use in bacterial cells. We provide TRX in a range of orientations, to meet all your cloning requirements. Please browse through our products using the ‘Browse Plasmids’ menu. For example we provide TRX at both N and C sides of the MCS, allowing versatile positioning of your gene, we sell it with and without enzyme cleavage sites (for simple removal of the tag, if required, after protein production). We also sell it in a range of configurations with other tags (such as hexahistidine) to give you maximum flexibility.

 

A typical TRX protein production is as follows:

  1. Harvest and lyse host cells with expressed TRX-tagged protein, and clarify by centrifugation

  2. TRX and many TRX fusion proteins are stable at 80°C, allowing initial purification by heat treatment to precipitate other proteins.

  3. Several types of affinity purification are possible, for example where an immobilized arsenical compound forms an adduct with the redox-sensitive vicinal dithiols present at the active site of thioredoxin. Pass the lysate slowly through an affinity column and wash carefully with buffer

  4. Elute by changing the pH and reducing potential of the buffer

  5. Purify by dialysis.

 

Our product range

Oxford Genetics provides a broad range of plasmids containing N-terminal and C-terminal TRX tags for mammalian, bacterial and yeast systems. In some plasmids the tags flank reporter genes, while in others they are positioned adjacent the MCS enabling you to insert your own gene in frame as required.

 

The plasmid structure shown has a TRX site downstream of the MCS, to be positioned at the C terminus of your gene of interest. The plasmid also provides a hexahistidine site, for improved purification, and both can be removed from the protein produced by Prescission cleavage of the 3C site, if required.

 

Our Plasmid Builder facility also provides a simple means for you to design an efficient strategy for any modifications or cloning manipulations to these plasmids. Finally, as always, although we have designed these plasmids for simple and efficient cloning, we are happy to undertake any cloning steps that you prefer to outsource. You can access all these options through the Plasmid Builder button below.

 

 

S-Tag Plasmid Vectors Information

S-tag properties and uses:

 

The S-tag is derived from the N terminus of RNase A. It is a 15 amino acid sequence that is used as an epitope tag and also as a solubility tag. The sequence is: Lys-Glu-Thr-Ala-Ala-Ala-Lys-Phe-Glu-Arg-Gln-His-Met-Asp-Ser. S-tag is suitable for use in many different cell types and has a molecular mass of 42.5 KDa.

 

Oxford Genetics provides a range of plasmid vectors containing S-tags for use in mammalian, bacterial and yeast cells. We provide S-tags in a range of orientations, to meet all your cloning requirements. Please browse through our products using the ‘Browse Plasmids’ menu. For example we provide S-tags at both N and C sides of the MCS, allowing versatile positioning of your gene, we sell it with and without enzyme cleavage sites (for simple removal of the tag, if required, after protein production), and we sell it in a range of configurations with other tags (such as hexahistidine) to give you maximum flexibility.

 

Purification of proteins using S-tags: the principle


  1. A typical S-tag-based purification is as follows:

  2. Harvest and lyse host cells with expressed S-tagged protein, and clarify by centrifugation

  3. Pass the lysate slowly through an affinity column containing anti-S-Tag antibodies and wash carefully with buffer.

  4. Elute the bound S-tagged protein and its binding partner(s) with a buffer containing high levels of free S-Tag peptide

 

Our product range

Oxford Genetics provides a broad range of plasmids containing N-terminal and C-terminal S tags for mammalian, bacterial and yeast systems. In some plasmids the tags flank reporter genes, while in others they are positioned adjacent the MCS enabling you to insert your own gene in frame as required.

 

The plasmid structure shown contains an S-Tag positioned downstream of the MCS, to be linked at the C terminal of your gene of interest. The plasmid also contains puromycin resistance driven by the ubiquitin promoter.

 

Our Plasmid Builder facility also provides a simple means for you to design an efficient strategy for any modifications or cloning manipulations to these plasmids. Finally, as always, although we have designed these plasmids for simple and efficient cloning, we are happy to undertake any cloning steps that you prefer to outsource. You can access all these options through the Plasmid Builder button below.

 

Simple plasmids containing S-Tag tags:

Multifunctional plasmids with S-Tag tags, protease sites for removal of tags after protein production, and many also contain additional functional tags:
 
 

C-Myc Tag Plasmid Vector Information

c-Myc tag properties and uses

 

c-Myc tags are decapeptide tags derived from the c-myc gene product that can be encoded within DNA sequences and be positioned at either end of the protein of interest. C-Myc tags can be used for affinity purification, co-precipitation and also as epitope tags for detection by western blotting.

 

Anti-c-Myc antibodies are efficient for detection in a range of formats. The c-Myc tag sequence, from N to C: N-Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu-C. It has a molecular weight of 1202 Da.

 

Immunoprecipitation of proteins using c-Myc tags: the principle

A typical c-Myc immunoprecipitation is as follows:

  1. Harvest and lyse host cells with expressed c-Myc-tagged protein, and clarify by centrifugation

  2. Incubate the lysate with anti-a-Myc antibodies and a slurry of Protein A-capture gel beads, centrifuge and wash.

  3. Elute the bound c-Myc-tagged protein with one of (a) a low pH glycine buffer, (b) SDS, (c) high pH urea buffer.

 

Our product range

Oxford Genetics provides a broad range of plasmids containing N-terminal and C-terminal c-Myc tags. In some plasmids the tags flank reporter genes, while in others they are positioned adjacent the MCS enabling you to insert your own gene in frame as required. The plasmid structure shown contains the c-Myc tag upstream of the MCS (where you can insert your gene of interest under the CMV promoter) and provides a thrombin cleavage site, allowing you to remove the c-Myc tag after production of your protein if you wish. It also contains the puromycin resistance gene under control of the ubiquitin promoter, for selection in mammalian cells, and kanamycin for selection in bacterial cells.

 

Our Plasmid Builder facility also provides a simple means for you to design an efficient strategy for any modifications or cloning manipulations to these plasmids. Finally, as always, although we have designed these plasmids for simple and efficient cloning, we are happy to undertake any cloning steps that you prefer to outsource. You can access all these options through the Plasmid Builder button below.

 

Simple plasmids containing c-Myc tags:

Multifunctional plasmids with c-Myc tags, protease sites for removal of tags after protein production, and many also contain additional functional tags:
 

Maltose Binding Protein Tags (MBP Tags) Plasmid Vectors Information

MBP tag properties and uses

The MBP tag is derived from the E. coli maltose/maltodextrin system and is widely used as both a solubility tag (notably for recombinant proteins grown in E. coli) and also as an affinity tag, since it binds to amylose columns. MBP has a molecular mass of 42.5 kDa.

 

Oxford Genetics provides a range of plasmid vectors containing MBP tags for use in mammalian, bacterial and yeast cells. We provide MBP in a range of orientations, to meet all your cloning requirements. Please browse through our products using the ‘Browse Plasmids’ menu. For example we provide MBP at both N and C sides of the MCS, allowing versatile positioning of your gene, we sell it with and without enzyme cleavage sites (for simple removal of the tag, if required, after protein production), and we sell it in a range of configurations with other tags (such as hexahistidine) to give you maximum flexibility.

 

Purification of proteins using MBP tags: the principle

 A typical MBP purification is as follows:

  1. Harvest and lyse host cells with expressed MBP-tagged protein, and clarify by centrifugation

  2. Pass the lysate slowly through an amylose column and wash carefully with buffer.

  3. Elute the bound MBP-tagged protein and its binding partner(s) with a buffer containing high levels of free maltose

  4. Purify by dialysis.

 

Our product range

Oxford Genetics provides a broad range of plasmids containing N-terminal and C-terminal MBP tags for mammalian, bacterial and yeast systems. In some plasmids the tags flank reporter genes, while in others they are positioned adjacent the MCS enabling you to insert your own gene in frame as required.

 

The plasmid structure shown includes an MBP tag positioned upstream of the MCS, for inclusion at the N terminus of your gene of interest, and an EKT cleavage site allowing simple removal of the MBP, if required, after protein production. Expression is regulated by the TEF1 promoter in yeast cells. The structure also contains URA3 for selection of transfected yeast cells in uracil-deficient media, and kanamycin resistance for selection in bacteria.

 

Our Plasmid Builder facility also provides a simple means for you to design an efficient strategy for any modifications or cloning manipulations to these plasmids. Finally, as always, although we have designed these plasmids for simple and efficient cloning, we are happy to undertake any cloning steps that you prefer to outsource. You can access all these options through the Plasmid Builder button below.

 

FLAG Tag Vector Plasmid Information

FLAG tag properties and uses

FLAG tags are octapeptide tags that can be encoded within DNA sequences and be positioned at either end of the protein of interest. FLAG tags can be used for affinity purification, co-precipitation and also for detection by western blotting. The FLAG tag is synthetic, and was designed to be small and hydrophilic, to exert minimal effects on the structure of proteins it is linked to.

 

Anti-FLAG antibodies are efficient for detection in a range of formats. The FLAG tag also contains an intrinsic enterokinase (EKT) cleavage site, for tag removal after it has fulfilled its purpose.

 

The FLAG tag sequence, from N to C: DYKDDDDK . It has a molecular weight of 1012 Da.

 

Immunoprecipitation of proteins using FLAG tags: the principle

A typical FLAG immunoprecipitation is as follows:

  1. Harvest and lyse host cells with expressed FLAG-tagged protein, and clarify by centrifugation

  2. Incubate the lysate with a slurry of FLAG-capture gel beads, centrifuge and wash carefully.

  3. Elute the bound FLAG-tagged protein and its binding partner(s) with a buffer containing the free FLAG-elution peptide DYKDDDDK.

 

Our product range

Oxford Genetics provides a broad range of plasmids containing N-terminal and C-terminal FLAG tags. In some plasmids the tags flank reporter genes, while in others they are positioned adjacent the MCS enabling you to insert your own gene in frame as required. The plasmid structure shown contains a FLAG tag upstream of the MCS, allowing you to position it simply at the N terminus of your gene of interest, and an enterokinase (EKT) site is also provided that allows simple removal of the FLAG tag from the purified protein if you so wish. In this case a BM40 (osteonectin) secretory tag is also included, to allow efficient secretion of your protein from the expressor cells.  Our online catalogue contains a broad range of plasmid structures, accessible via the buttons below.

 

Our Plasmid Builder facility also provides a simple means for you to design an efficient strategy for any modifications or cloning manipulations to these plasmids. Finally, as always, although we have designed these plasmids for simple and efficient cloning, we are happy to undertake any cloning steps that you prefer to outsource. You can access all these options through the Plasmid Builder button below.

 

 

Protein Tag Guide

Protein Tag Guide

Protein tags can be used for a variety of protein applications, including:

  • Protein secretion
  • Affinity purification
  • Epitope detection for western blots, FACS or immunohistochemistry
  • Chromatography
  • Increasing solubility
  • Reporter gene fusions to measure protein levels and activity


Oxford Genetics Ltd provides the largest collection of protein tags in the world, with over 1500 peptide tag variant plasmids for most research purposes. We can also easily create any new tag you need on request if we haven’t got the combination you need already. When choosing which protein tag to use it is important to consider your required application. Some tags are much more efficient than others for certain applications and the table below may help you decide which to use.

Typically, affinity tags are useful for protein purification, whilst solubility tags can be useful for increasing yields (but often also for purification). Epitope tags are typically useful for protein detection, but can also be used for purification but this is generally more expensive than using affinity resins because they require large amounts of antibody for purification.

As general rule you can normally find antibodies against most of the tags available, so technically they can all be used for detection and/or purification, but using certain methods for some tags can be much more expensive and some can produce less pure protein preparations.

 

Affinity Tags                                             

Tag Name Size Level of Affinity  Common Resins Relative Cost
Deca-Histidine (10His)  10 *** Talon or Ni-NTA *
Hexa-Histidine (6His) 6 *** Talon or Ni-NTA *
Strep 8 *** Strep-Tactin–Sepharose **
Glutathione-S-Transferase (GST) 218 ** GSH–Sepharose *
Maltose Binding Protein (MBP) 396 * Amylose *

 

 

Epitope Tags   

 

Tag Name Size Method of Purification 

Cost (antibody specific)

Sequence
FLAG 8 Antibody Purification *** DYKDDDDK
C-Myc 10 Antibody Purification *** EQKLISEEDL
HA 9 Antibody Purification *** YPYDVPDYA
T7 11 Antibody Purification *** MASMTGGQQMG
V5 14 Antibody Purification *** GKPIPNPLLGLDST
S-tag 15 Antibody Purification *** KETAAAKFERQHMDS
Glu-Glu 9 Antibody Purification *** EYMPME
HSV 12 Antibody Purification *** QPELAPEDPEDC
VSV 11 Antibody Purification *** YTDIEMNRLGK
E-Tag 13 Antibody Purification *** GAPVPYPDPLEPR
Strep Tag 8 Antibody Purification *** WSHPQFEK

 

Reporter Tags            

 

Reporter Name

Ease of

Detection

Reporter Type Location Length (AAs)
FireFly Luciferase (FLuc) *** Luminescent Cystosol 549
Renilla Luciferase (RLuc) ** Luminescent Cystosol 311
iLumena Luciferase *** Luminescent Secreted 211
Beta Galactosidase (BetaGal) ** Colourimetric Cystosol 1047
Secreted Alkaline Phosphatase (SEAP) ** Colourimetric Secreted 520
Chloramphenicol Acetyl Transferase (CAT) * Colourimetric Cystosol 218
daGFP *** Fluorescent Cystosol 235
KrYFP *** Fluorescent Cystosol 235
FrCFP * Fluorescent Cystosol 235

 

If you require any further information regarding our product range then please contact us for more information.