A polyhistidine-tag is an amino acid motif in proteins that consists of at least six histidine (His) residues, often at the N- or C-terminus of the protein. It is also known as hexa histidine-tag, 6xHis-tag, His6 tag and by the trademarked name His-tag. Polyhistidine-tags are often used for affinity purification of polyhistidine-tagged recombinant proteins expressed in Escherichia coli and other prokaryotic expression systems. To purchase this antibody, use the following link: http://www.stjohnslabs.com/his-tag-antibody-p-95792?filter_name=STJ90106

1. HOW DID OUR
ANTIBODY PERFORM?
CUSTOMER REVIEW: His-tag Polyclonal Antibody (STJ90106)

2. WHAT WAS USED?
Primary antibody: Product Catalog No: STJ90106. His-tag
polyclonal antibody. 100µl
Provider: St John's Laboratory
Dilution ratio: 1:1000
Secondary antibody:
HRP goat anti-rabbit (provided by BPI)
Reference antibody:
Actin (provided by BPI)Figure 1. Lane1: HeLa; Lane 2: Jurkat

3. WHAT WAS THE PROTOCOL?
Antibody: STJ90106. His-tag antibody
Method for validation: Western Blot
Materials for validation: HeLa cell line, Jurkat
cell line
Treatment of materials: When cells were cultured
to 70%-80% plating density, lysis buffer with
protease inhibitors (Sigma) and equal volume of
2X loading buffer were added for 10 min water
bath at 100˚C.Figure 1. Lane1: HeLa; Lane 2: Jurkat

4. WHAT WAS THE PROTOCOL?
Gel electrophoresis: 10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer: 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant
voltage 100 V 60 min.
Blocking: 1X TBST with 5% skimmed milk powder for 2 hr.
Primary antibody probing: primary antibody was added to the solution of 1X
TBST with 5% skimmed milk powder, 4˚C for overnight.
Membrane wash: 1X TBST wash for 3 times.
Secondary antibody probing: secondary antibody was added to the solution of
1X TBST with 5% skimmed milk powder with 1:10000 dilution ratio, for 1.5 hr.
Membrane wash: 1X TBST wash for 3 times.
Visualization: ECL visualization for 5 min (the visualization time was
determined by actual result. It was ensured that the visualization time was
identical for the same antigen).
Figure 1. Lane1: HeLa; Lane 2: Jurkat

5. WHAT WAS THE PROTOCOL?
Gel electrophoresis information：10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information：0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. Antigen
Loading
amount
Primary
antibody
Primary
antibody
dilution
ratio
Secondary
antibody
dilution ratio
Target band
KD
Visualization
time
1 HeLa lysate 10 µg STJ90106.
His-tag
antibody
1:1000
1:10000 ~20KD 20S
2
Jurkat
lysate
10 µg 1:10000 ~20KD 20S
Figure 1. Lane1: HeLa; Lane 2: Jurkat

6. WHAT DID THE CUSTOMER THINK?
Testing results were provided by BPI laboratory. They
stated that “In HeLa and Jurkat cell line, there were target
bands detected by STJ90106 His-tag Antibody”.

7. “WE INVITE YOU TO TEST OUR PRODUCTS”
Our aim is to not only connect our customers
with the best product, but to also give them
the opportunity to test the product’s
performance risk free.

8. Contact Us: info@stjohnslabs.com
1. Choose 5 Trial Size Samples from our free sample list
2. Complete the free sample application: http://www.stjohnslabs.com/free-sample
3. Receive your antibody
4. Submit your review
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