Revision 3
#2366
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877-616-CELL (2355)
877-678-TECH (8324)
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, W-S, IP
Reactivity:
All
Sensitivity:
Transfected Only
Source/Isotype:
Mouse IgG1
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Simple Western™ | 1:10 - 1:50 |
| Immunoprecipitation | 1:200 |
Storage
Specificity/Sensitivity
Source / Purification
Background
A variety of plasmids contain DNA that encodes an amino-terminal tag cosisting of six histidine (6xHis) residues followed by an extended multiple cloning site. The 6xHis tag on the expressed recombinant proteins allows for efficient coupling to Ni2+ affinity resins and purification by single step chromatography (1).
As is the case with other protein tag systems (2), this polyhistidine tag can often be cleaved at sites recognized by proteases such as thrombin and enterokinases to isolate the protein of interest (1).
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation
Cross-Reactivity Key
All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
限制使用
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Revision 3
#2366
His-Tag (27E8) Mouse mAb
Simple Western™ analysis of lysates (0.1 mg/mL) from COS-7 cells, transfected with mAkt1-Myc/His (C-terminal Myc/His tag), using His-Tag (27E8) Mouse mAb #2366. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
