|
TAG (Abbreviation) |
Recommended Resin |
Vendor (with link) |
Manuf. Stated Resin Capacity |
Pre‐elution wash* |
MS‐Compatible Elution* |
Cleanup |
|
Green Fluorescent Protein (GFP) |
GFP‐Trap_A** GFP‐Trap_M** |
~2 ug GFP per 10 uL resin. |
0.5% to 1% NP40 in PBS, then 50 mM Ammonium Bicarbonate (3x) |
0.25% Rapigest SF, 50mM AmBic, Boil Or Supply resin for direct on‐resin digestion (consult DPCF for preferred approach) |
None required |
|
|
Glutathione S‐ Transferase (GST) |
Glutathione Sepharose 4B** (GE Healthcare; 17‐ 0756‐01) |
Not given. |
0.5% to 1% NP40 in PBS, then 50 mM Ammonium Bicarbonate (3x) |
Excess glutathione in 50mM AmBic, no detergents. Or Supply resin for direct on‐resin digestion (consult DPCF for preferred approach) |
If >150uL, use Amicon 4 10kDa MWCO. For less, use Zebaspin desalt gel filtration columns. |
|
|
FLAG |
ANTI‐FLAG M2 Affinity Gel (A2220) |
>0.6 mg/mL |
0.5% to 1% NP40 in PBS, then 50 mM Ammonium Bicarbonate (3x) |
200 ug/mL FLAG peptide in 50mM AmBic, no detergents. |
If >150uL, use 10kDa MWCO. For less, Zebaspin desalt gel filtration columns. |
|
MYC |
ANTI‐MYC Affinity Agarose (A7470) |
2 nmol/mL |
0.5% to 1% NP40 in PBS, then 50 mM Ammonium Bicarbonate (3x) |
0.25% Rapigest in 50 mM AmBic, with boiling. |
None required. Immediately compatible for LC‐ MS/MS. |
|
|
Poly‐histidine (6xHis) |
HisPur Cobalt Resin (Pierce 89964)** Ni‐NTA Agarose (Quiagen 30210)** |
>10 mg/ml His‐ tagged protein (HisPur) |
0.5% to 1% NP40 in PBS, then 50 mM Ammonium Bicarbonate (3x) |
10mM – 200 mM imidazole in 50mM AmBic, no detergents; or on‐resin digestion. |
If >150uL, use Amicon 4 10kDa MWCO. For less, use Zebaspin desalt gel filtration columns. |
|
|
Biotin (peptide pulldown) |
Monomeric avidin |
1.2 mg biotinylated BSA/mL resin |
0.1% SDS in PBS at RT (30 min), then 50mM AmBic (3x) |
0.4/30/69.6 v/v/v TFA/MeCN/ water |
None required. Immediately compatible for LC‐ MS/MS. |
|
|
Biotin (protein pulldown) |
Streptavidin Agarose |
Up to 10 mg biotinylated BSA/mL resin |
0.1% SDS in PBS at RT (30 min), then 50mM AmBic (3x) |
200 ug/mL Biotin in PBS, or boil in 0.5% Rapigest/50 mM AmBic |
Zebaspin x2 for biotin elution. None required for Rapigest Elution |
|
|
HA Tag |
Pierce HA Tag IP Kit/Resin (Pierce 26181) |
60 nmol HA‐ tagged protein per mL slurry. |
0.5% to 1% NP40 in PBS, then 50 mM Ammonium Bicarbonate (3x) |
0.1 M Glycine (pH 2.8), or 0.25% Rapigest SF with boiling. |
Zebaspin or Amicon MWCO for Glycine elution. None required for Rapigest elution. |