PKM2 Antibody

Images

 
Western Blot: PKM2 Antibody [NBP1-48308] - Analysis of PKM2 in MCF7 whole cell lysates.
Immunocytochemistry/ Immunofluorescence: PKM2 Antibody [NBP1-48308] - IF analysis of PKM2 in frozen sections of rat glioblastoma tissue. Image courtsey of anonymous customer review.
Immunohistochemistry: PKM2 Antibody [NBP1-48308] - IHC staining of PKM2 in mouse liver tissue.
Western Blot: PKM2 Antibody [NBP1-48308] - Analysis of HeLa lysates using NBP1-48308. Image courtesy of Gregg Semenza - publication number 21620138 (PMID).
Immunocytochemistry/ Immunofluorescence: PKM2 Antibody [NBP1-48308] - Immunocytochemical analysis of PKM2 in HeLa cells.
Simple Western: PKM2 Antibody [NBP1-48308] - Simple Western lane view shows a specific band for PKM2 in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230kDa separation ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, BvSpecies Glossary
Applications WB, Simple Western, ICC/IF, IHC, IHC-P
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.41 mg/ml

PKM2 Antibody Summary

Immunogen
A synthetic peptide made to the internal region of human PKM2 protein (within residues 350-450). [Swiss-Prot# P14618]
Localization
Cytoplasm. Nucleus. Note: Translocates to the nucleus in response to different apoptotic stimuli. Nuclear translocation is sufficient to induce cell death that is caspase independent, isoform-specific and independent of its enzymatic actvity.
Predicted Species
Bovine (95%). Backed by our 100% Guarantee.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
PKM
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 0.5 ug/ml
  • Simple Western 1:12.5
  • Immunocytochemistry/Immunofluorescence 1:200
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Paraffin 1:100
Application Notes
This PKM2 antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry on paraffin-embedded sections and Western blot, where a band is seen at approx. 60 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Theoretical MW
60 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Positive Control
HeLa Lysate (NB800-PC1)
MCF-7 Lysate (NB820-59465)
Control Peptide
PKM2 Peptide (NBP1-48308PEP)
Reviewed Applications
Read 3 Reviews rated 5
using
NBP1-48308 in the following applications:

Publications
Read Publications using
NBP1-48308 in the following applications:

  • 2 publications
  • 1 publication
  • IP
    1 publication
  • WB
    8 publications

Reactivity Notes

Human, mouse and rat. Immunogen sequence has 95% identity to bovine.

Packaging, Storage & Formulations

Storage
Store at -20C. Avoid freeze-thaw cycles.
Buffer
PBS and 30% Glycerol
Preservative
0.1% Sodium Azide
Concentration
1.41 mg/ml
Purity
Immunogen affinity purified

Alternate Names for PKM2 Antibody

  • CTHBP
  • Cytosolic thyroid hormone-binding protein
  • MGC3932
  • OIP3
  • OIP-3
  • OIP3EC 2.7.1.40
  • Opa-interacting protein 3
  • p58
  • PK, muscle type
  • PK2
  • PK3
  • PK3PKM
  • PKM2
  • Pyruvate kinase 2/3
  • pyruvate kinase isozymes M1/M2
  • Pyruvate kinase muscle isozyme
  • pyruvate kinase, muscle
  • TCB
  • THBP1
  • THBP1p58
  • Thyroid hormone-binding protein 1
  • thyroid hormone-binding protein, cytosolic
  • Tumor M2-PK

Background

PKM2 (pyruvate kinase isozymes M1/M2), an isoform of pyruvate kinase, is a glycolytic enzyme that catalyzes the transfer of a phosphoryl group from phosphoenolpyruvate (PEP) to ADP, generating ATP and is expressed in proliferating cells, such as embryonic stem cells, embryonic carcinoma cells, as well as cancer cells. Other three pyruvate kinases includes PKL (liver and kidney), PKR (erythrocytes) and PKM1, (adult muscle, brain, bladder, and fibroblasts) which exist as stable tetramers, whereas, PKM2 subunits form tetramers as well as dimers, the dimeric form with higher Km value for PEP substrate stays inactive at physiological concentrations of PEP. The ratio between highly active tetrameric form and nearly inactive dimeric form determines whether glucose carbons are channeled to biosynthetic processes or used for glycolytic ATP production. Predominately localized in the cytoplasm, PKM2 translocate to nucleus upon apoptotic stimuli and this translocation mediated cell death is caspase independent, isoform-specific and independent of its enzymatic activity.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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NBP1-48308
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Publications for PKM2 Antibody (NBP1-48308)(11)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 4 applications: ICC/IF, IHC-P, IP, WB.


Filter By Application
ICC/IF
(2)
IHC-P
(1)
IP
(1)
WB
(8)
All Applications
Filter By Species
Human
(8)
Mouse
(1)
All Species
Showing Publications 1 - 10 of 11. Show All 11 Publications.
Publications using NBP1-48308 Applications Species
Minami K, Taniguchi K, Sugito N et al. MiR-145 negatively regulates Warburg effect by silencing KLF4 and PTBP1 in bladder cancer cells. Oncotarget. 2017 May 16 [PMID: 28380435] (WB, Human) WB Human
Takai T, Yoshikawa Y, Inamoto T et al. A Novel Combination RNAi toward Warburg Effect by Replacement with miR-145 and Silencing of PTBP1 Induces Apoptotic Cell Death in Bladder Cancer Cells Int J Mol Sci Jan 17 2017 [PMID: 28106737] (WB, Human) WB Human
Sugiyama T, Taniguchi K, Matsuhashi N et al. MiR-133b inhibits growth of human gastric cancer cells by silencing pyruvate kinase muscle-splicer polypyrimidine tract-binding protein 1. Cancer Sci. Dec 1 2016 [PMID: 27696637] (WB, Human) WB Human
Christensen DR, Calder PC, Houghton FD. GLUT3 and PKM2 regulate OCT4 expression and support the hypoxic culture of human embryonic stem cells. Sci Rep. 2015 Dec 07 [PMID: 26639784] (WB, ICC/IF, Human) WB, ICC/IF Human
Taniguchi K, Sugito N, Kumazaki M et al. Positive feedback of DDX6/c-Myc/PTB1 regulated by miR-124 contributes to maintenance of the Warburg effect in colon cancer cells Biochim. Biophys. Acta. 2015 Jul 02 [PMID: 26144048] (WB, Human)

Details:
PKM2 antibody was used for WB assay on lysates of human colon cancer cells (DLD-1 cells and WiDr cells ) transfected with miR-124 or siR-PTB1 (Fig. 3B and 3C). WB was also performed on lysates after the transfection of colon cancer cells with siR-c-Myc or siR-DDX6 (Fig. 4A qand 4B), and on DLD-1 cells /WiDr cells that were subjected to PKM1 and/or PKM2 knockdown using siRNAPKM1 and siRNAPKM2 (Fig. 5B and 5E).
WB Human
Taniguchi K, Ito Y, Sugito N et al. Organ-specific PTB1-associated microRNAs determine expression of pyruvate kinase isoforms Sci Rep 2015 Feb 27 [PMID: 25721733] (WB, ICC/IF, IHC-P, Mouse, Human) WB, ICC/IF, IHC-P Mouse, Human
Shinohara H, Taniguchi K, Kumazaki M et al. Anti-cancer fatty-acid derivative induces autophagic cell death through modulation of PKM isoform expression profile mediated by bcr-abl in chronic myeloid leukemia Cancer Lett. 2015 Jan 30 [PMID: 25644089] (WB, Human)

Details:
PKM2 antibody used for WB on lysates of human leukemia cell lines K562 and KCL-22 cells which were subjected to AIC-47/a fatty-acid derivative treatment for 72?hours (Fig.?3B). WB also performed on the lysates of K562 and KCL-22 cells at 72?h after transfection of the cells with control RNA or siRNAs against c-Myc, PTB1 or PKM2 (Fig.?3C, 3D and 3E ), and with control RNA or bcr-abl-specific siRNA /siR-bcr-abl (Fig.?4). PKM1 WB also performed on lysates of K562 cells transfected with control siRNA or siRNA-STAT3 (Figure S1A) and cells treated or not with Rapamycin (Figure S2).
WB Human
Bruns Dr, Brown Rd, Stenmark Kr et al. Mitochondrial Integrity in a neonatal Bovine Model of Right Ventricular Dysfunction. Am. J. Physiol. Lung Cell Mol. Physiol. 2014 Nov 21 [PMID: 25416385]
Lindsay Kj, Du J, Sloat Sr et al. Pyruvate kinase and aspartate-glutamate carrier distributions reveal key metabolic links between neurons and glia in retina. Proc. natl. Acad. Sci. U.S.A. 2014 Oct 28 [PMID: 25313047]
Sasaki M, Fujimoto S, Sato Y et al. Reduction of reactive oxygen species ameliorates metabolism-secretion coupling in islets of diabetic GK rats by suppressing lactate overproduction. Diabetes 2013 Jan 24 [PMID: 23349483]
Show All 11 Publications.

Reviews for PKM2 Antibody (NBP1-48308) (3) 53

Average Rating: 5
(Based on 3 reviews)
We have 3 reviews tested in 2 species: Human, Rat.
We have 3 reviews tested in 3 application: ICC, WB, IF.

Reviews using NBP1-48308:
Filter by Applications
ICC
(1)
WB
(1)
IF
(1)
All Applications
Filter by Species
Human
(2)
Rat
(1)
All Species
Images Ratings Applications Species Date Details
 
reviewed by:
Sachin Puri
ICC Human 06/15/2017
View

Summary

ApplicationImmunocytochemistry
Sample TestedHuman lung cancer cell lines
SpeciesHuman
Western Blot PKM2 NBP1-48308
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reviewed by:
Kelsey Stockbridge
WB Human 02/17/2011
View

Summary

ApplicationWestern Blot
Sample TestedHeLa cells
SpeciesHuman

Blocking

Blocking DetailsBlocking Buffer: 5% milk in TBST, Blocking Time: 1 hour, Blocking Temp: 25 degrees Celsius

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:1000, Incubation Dilution Buffer: 5% Milk in TBST, Incubation Time: overnight, Incubation Temp: 4C

Secondary Antibody

Secondary DescriptionSecondary Ab: Donkey anti-Rabbit IgG HRP, Secondary Ab Dilution Ratio: 1:5000

Details

Detection NotesDetection Method: ECL Plus, Specific Bands: 58 kDa
Immunofluorescence PKM2 NBP1-48308
Enlarge
reviewed by:
Jian Zhang
IF Rat 10/15/2010
View

Summary

ApplicationImmunofluorescence
Sample TestedRat glioblastoma frozen section
SpeciesRat
Comments"I have tried this PKM2 antibody for both western blots and for immunofluorescence, and the results were very good.</p><p>Western blots went very well on human, mouse, and rat cell lines (I have tried 293T, HeLa, 3T3, C2C12, and C6 cell lines). The specific band (~58KD) gives very bright signal, with little background (I used 1:500 dilution). Our later experiments confirmed that this antibody is specific for the M2 isoform of Pyruvate kinase (PKM2), not the M1 isoform (PKM1). </p><p>The best thing about this antibody is that it works well for immunofluorescence staining of frozen sections of tumor samples (rat glioblastoma). The image I provided shows that PKM2 expresses predominantly in the cytoplasm of hnRNPA1+ cells (red nucleus), not hnRNPA- cells (blue nucleus). That is exactly what we expected. This result further confirms that the antibody is specific for PKM2, not PKM1 (which is expected to be present in hnRNPA1- cells). I used 1:200 dilution (at 4 C overnight) for immunofluorescence.</p><p>"

Blocking

Blocking DetailsBlocking Buffer: PBS + 0.1% Tween 20 + 20% normal goat serum, Blocking Time: 30 minutes, Blocking Temp: room temperature

Primary Anitbody

Dilution RatioIncubation Dilution Buffer:PBS + 0.1% Tween 20+20% norm goat serum, Dilution Ratio: 1:200, Incubation Time:overnight, Temp:4C

Secondary Antibody

Secondary DescriptionSecondary Ab: Alexa Fluor 488 goat anti-rabbit IgG, Secondary Ab Dilution: 1:1000
Secondary Manufacturer Cat#A11034

Details

Detection NotesDetection Method: Fluorescence at 488nm, Pos Ctrl: hnRNPA1 Ab, Neg Ctrl: Blocking buffer only (instead of PKM2 ab)
Fixation DetailsSample Preparation: PFA fixed

Comments

Comments"I have tried this PKM2 antibody for both western blots and for immunofluorescence, and the results were very good.</p><p>Western blots went very well on human, mouse, and rat cell lines (I have tried 293T, HeLa, 3T3, C2C12, and C6 cell lines). The specific band (~58KD) gives very bright signal, with little background (I used 1:500 dilution). Our later experiments confirmed that this antibody is specific for the M2 isoform of Pyruvate kinase (PKM2), not the M1 isoform (PKM1). </p><p>The best thing about this antibody is that it works well for immunofluorescence staining of frozen sections of tumor samples (rat glioblastoma). The image I provided shows that PKM2 expresses predominantly in the cytoplasm of hnRNPA1+ cells (red nucleus), not hnRNPA- cells (blue nucleus). That is exactly what we expected. This result further confirms that the antibody is specific for PKM2, not PKM1 (which is expected to be present in hnRNPA1- cells). I used 1:200 dilution (at 4 C overnight) for immunofluorescence.</p><p>"

Product General Protocols

View specific protocols for PKM2 Antibody (NBP1-48308): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for PKM2 Antibody (NBP1-48308). (Showing 1 - 1 of 1 FAQs).

  1. I am wondering if this antibody can be used for IP or ChIP? Have you ever tested it in either of these applications?
    • We have not yet tested this antibody in ChIP or IP. However, if you would like to test this antibody in either of these applications and share your results with us, you would become eligible for our Innovators Reward Program.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NBP1-48308AF405
Alexa Fluor 488 Labeled NBP1-48308AF488
Alexa Fluor 647 Labeled NBP1-48308AF647
Alexa Fluor 700 Labeled NBP1-48308AF700
Biotin Labeled NBP1-48308B
DyLight 405 Labeled NBP1-48308V
DyLight 488 Labeled NBP1-48308G
DyLight 550 Labeled NBP1-48308R
DyLight 650 Labeled NBP1-48308C
FITC Labeled NBP1-48308F
HRP Labeled NBP1-48308H

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MAPK3/ERK1 - A signal transduction pathway with roles in development and disease
Mitogen-activated protein kinases (MAPKs) are important signaling proteins needed to transmit and relay extracellular stimuli and to illicit intracellular responses (1). The MAPK family of proteins are serine/threonine kinases that are able to phos...  Read full blog post.

5 Stars for the PKM2 Antibody
Novus' Pyruvate Kinase M2 antibody (cat # NBP1-48308) has received some glowing product reviews and customer feedback lately. One satisfied customer wrote, "The best thing about this antibody is that it works well for immunofluorescence staining of fr...  Read full blog post.

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Recent Reviews

5
5
3
4
0
3
0
2
0
1
0

 
Sachin Puri
06/15/2017
Application: ICC
Species: Human

Kelsey Stockbridge
02/17/2011
Application: WB
Species: Human

Jian Zhang
10/15/2010
Application: IF
Species: Rat

Bioinformatics

Gene Symbol PKM
Entrez
Uniprot