NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. To purchase this antibody, use the following link: http://www.stjohnslabs.com/nfkb-p65-antibody-p-93367?filter_name=STJ94468

1. HOW DID OUR
ANTIBODY PERFORM?
CUSTOMER REVIEW: NFκB-p65 Polyclonal Antibody (STJ94468)

2. WHAT WAS USED?
Primary antibody: STJ94468. NFκB-p65 Polyclonal
Antibody. 100µl
Provider: St John's Laboratory
Dilution ratio: 1:1000
Application: Western Blot.
Image: In HeLa cell line, there was a target band detected
by STJ94468. NFκB-p65 Polyclonal Antibody.In A549 cell
line, there were no target bands detected by STJ94468
NFκB-p65 Polyclonal Antibody.

3. WHAT WAS THE PROTOCOL?
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.
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).

4. 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 STJ94468.
NFκB-p65
Polyclonal
Antibody
1:1000
1:10000 65KD 30S
2 A549 lysate 10 µg 1:10000 65KD 30S

5. WHAT DID THE CUSTOMER THINK?
Testing results were provided by BPI laboratory. They
stated that “In HeLa cell line, there was a target band
detected by STJ94468. NFκB-p65 Polyclonal Antibody. In
A549 cell line, there were no target bands detected by
STJ94468 NFκB-p65 Polyclonal Antibody”.

6. “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.

7. 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
5. Receive your 30% off your next order
Get Your Free Samples in 5 Easy Steps

8. Follow Us for Weekly Independent
Validation Reviews
@StJohnsLabsSt John's Laboratory Ltd
St John's Laboratory Ltd St John's Laboratory Ltd
http://www.slideshare.net/stjohnslabs