Pyruvate kinase isozymes M1/M2 (PKM1/M2), also known as pyruvate kinase muscle isozyme (PKM), pyruvate kinase type K, cytosolic thyroid hormone-binding protein (CTHBP), thyroid hormone-binding protein 1 (THBP1), or opa-interacting protein 3 (OIP3), is an enzyme that in humans is encoded by the PKM2 gene. Is a glycolytic enzyme that catalyzes the transfer of a phosphoryl group from phosphoenolpyruvate (PEP) to ADP, generating ATP. It plays a general role in caspase independent cell death of tumor cells. The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. Anti-PMK2 - http://www.stjohnslabs.com/pkm2-antibody-p-93954?filter_name=Stj95142 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

1. Result
HEK293 produced strong signal, In U87, HeLa and Jurkat cell lysates target bands were not detected. PKM2 antibody (STJ95142). (Figure:
Lane1: HEK293; Lane 2: U87; Lane 3:HeLa; Lane 4: Jurkat).
Cell Line:
HEK293,U87 cell lysate, HeLa
cell lysate, Jurkat
Method of validation: Western Blot
Primary Antibody: STJ95142 PKM2 antibody
Secondary Antibody
Polyclonal Goat anti Rabbit- HRP
Conjugated (Dako)
Reference antibody: GAPDH (abcam)
Dilution ratio: 1:1000
Protocol
Treatment of materials:
Cell pellets were lysed in RIPA lysis buffer with protease inhibitor cocktail (Roche) and equal volume of 2X loading buffer (Sigma)
were heated at 95˚C for 10 min in a heat block
E.g. Gel electrophoresis: 12% polyacrylamide gel, constant voltage 150 V, 60 min.
Transfer: 0.45 µm NC membrane, 1 piece of gel (wet transfer), constant voltage 40 V 120 min.
Blocking: 1X TBST with 5% skimmed milk powder over night.
Primary antibody probing: primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder, incubation on
the shaker, room temperature for 1hr
Membrane wash: 1X TBST wash for 3 times with each time 5minutes incubation on the shaker.
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 with each time 5minutes incubation on the shaker and final wash with deionized wa-
ter.
Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that the visualiza-
tion time was identical for the same antigen).
Antibody Customer Review:
STJ95142 Anti-PKM2 antibody
Antibody Specificity:
Antibody Rating:
Gel electrophoresis information：12% polyacrylamide gel, constant voltage 185 V, 60 min.
Transfer information：0.45 µm NC membrane (Bio Rad), 1 piece of gel (wet transfer), constant voltage 40 V 120 min.
No. Antigen
Loading
amount
Primary anti-
body
Primary
antibody
dilution
ratio
Secondary
antibody dilu-
tion ratio
Target band KD Visualization time
1
HEK293 cell
lysate
18µg(total
protein)
STJ95142.
PKM2 anti-
body
1:1000
1:5000 ~58KD 120S
2
U87 cell
lysate
18µg (total
protein)
1:5000 ~58KD 120S
3 HeLa Cell lysate
18µg (total
protein)
STJ95142
PKM2 anti-
body
1:1000 1:5000 ~58KD 120S
4
Jurkat Cell ly-
sate
18µg (total
protein)
STJ95142
PKM2 anti-
body
1:1000 1:5000 ~58KD 120S
GAPDH
Marker 1 2 3 4