PKM2 Antibody - BSA Free

Novus Biologicals | Catalog # NBP1-48308

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Bovine

Cited:

Human, Mouse, Bovine

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunoblotting, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Knockdown Validated

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all PKM2 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to the internal region of human PKM2 protein (within residues 350-450). [Swiss-Prot: P14618]

Reactivity Notes

Bovine reactivity reported in scientific literature (PMID: 25416385).

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 activity.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

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.

Scientific Data Images for PKM2 Antibody - BSA Free

Western Blot: PKM2 AntibodyBSA Free [NBP1-48308]

Western Blot: PKM2 AntibodyBSA Free [NBP1-48308]

PKM2-Antibody---BSA-Free-Western-Blot-NBP1-48308-img0018.jpg
Immunocytochemistry/ Immunofluorescence: PKM2 Antibody - BSA Free [NBP1-48308]

Immunocytochemistry/ Immunofluorescence: PKM2 Antibody - BSA Free [NBP1-48308]

Immunocytochemistry/Immunofluorescence: PKM2 Antibody - BSA Free [NBP1-48308] - Analysis of PKM2 in frozen sections of rat glioblastoma tissue. Image courtsey of anonymous customer review.
Immunohistochemistry: PKM2 Antibody - BSA Free [NBP1-48308]

Immunohistochemistry: PKM2 Antibody - BSA Free [NBP1-48308]

Immunohistochemistry: PKM2 Antibody - BSA Free [NBP1-48308] - Staining of PKM2 in mouse liver tissue.
Flow Cytometry: PKM2 Antibody - BSA Free [NBP1-48308]

Flow Cytometry: PKM2 Antibody - BSA Free [NBP1-48308]

Flow Cytometry: PKM2 Antibody - BSA Free [NBP1-48308] - An intracellular stain was performed on Neuro2a cells with PKM2 Antibody NBP1-48308AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.
Western Blot: PKM2 AntibodyBSA Free [NBP1-48308]

Western Blot: PKM2 AntibodyBSA Free [NBP1-48308]

Western Blot: PKM2 Antibody - BSA Free [NBP1-48308] - Analysis of HeLa lysates using NBP1-48308. Image courtesy of Gregg Semenza, (PMID: 21620138).
Western Blot: PKM2 AntibodyBSA Free [NBP1-48308]

Western Blot: PKM2 AntibodyBSA Free [NBP1-48308]

Western Blot: PKM2 Antibody - BSA Free [NBP1-48308] - Analysis of PKM2 in MCF7 whole cell lysates.
Western Blot: PKM2 AntibodyBSA Free [NBP1-48308]

Western Blot: PKM2 AntibodyBSA Free [NBP1-48308]

PKM2-Antibody---BSA-Free-Western-Blot-NBP1-48308-img0016.jpg
Western Blot: PKM2 AntibodyBSA Free [NBP1-48308]

Western Blot: PKM2 AntibodyBSA Free [NBP1-48308]

PKM2-Antibody---BSA-Free-Western-Blot-NBP1-48308-img0017.jpg
Immunocytochemistry/ Immunofluorescence: PKM2 Antibody - BSA Free [NBP1-48308]

Immunocytochemistry/ Immunofluorescence: PKM2 Antibody - BSA Free [NBP1-48308]

Immunocytochemistry/Immunofluorescence: PKM2 Antibody - BSA Free [NBP1-48308] - Analysis of PKM2 in HeLa cells.
Immunohistochemistry: PKM2 Antibody - BSA Free [NBP1-48308]

Immunohistochemistry: PKM2 Antibody - BSA Free [NBP1-48308]

PKM2 Antibody - BSA Free-Immunohistochemistry-NBP1-48308-img0020.jpg
Flow Cytometry: PKM2 Antibody - BSA Free [NBP1-48308]

Flow Cytometry: PKM2 Antibody - BSA Free [NBP1-48308]

Flow Cytometry: PKM2 Antibody - BSA Free [NBP1-48308] - An intracellular stain was performed on HeLa cells with PKM2 Antibody NBP1-48308AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.
Simple Western: PKM2 AntibodyBSA Free [NBP1-48308]

Simple Western: PKM2 AntibodyBSA Free [NBP1-48308]

Simple Western: PKM2 Antibody - BSA Free [NBP1-48308] - 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 system. * Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
PKM2 Antibody - BSA Free

Immunohistochemistry-Paraffin: PKM2 Antibody - BSA Free [NBP1-48308] -

Expression of PKM1 and PKM2 in clinical colorectal cancer samples.(a) The protein expression of PKM1 and PKM2 in clinical specimens of cancer tumor (T) and the adjacent normal tissues (N) is shown. PKM1 and PKM2 were detected by Western blotting in under the same experimental conditions at the same time. The full-length blots are presented in Supplementary Figure S3b. (b–d) Immunohistochemical staining of normal colon tissue adjacent to tumor tissue of case 10. Results of H&E staining (b), staining with anti-PKM1 antibody (c), and staining with anti-PKM2 (d) are shown. The boxed regions in “c" and “d" are enlarged in the images below. (e–h) Immunohistochemical staining of clinical colorectal cancer tissue specimen of representative case 3. H&E-stained section with normal tissue (upper right corner) neighboring the tumor area in the section is shown (e), along with the same section stained with anti-PKM2 antibody (f). Enlarged views of boxed areas in “f" show normal colorectal crypt in mucosa (g) and tumor area (h) stained with anti-PKM2 antibody. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25721733), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for PKM2 Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

5 ug/ml

Immunoblotting

reported in scientific literature (PMID 25416385)

Immunocytochemistry/ Immunofluorescence

1:200

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Immunoprecipitation

reported in scientific literature (PMID 21620138)

Simple Western

1:12.5

Western Blot

0.5 ug/ml
Application Notes
In Western blot, 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.
See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 0.1 mg/mL, separated by Size, antibody dilution of 1:12.5, apparent MW was 74 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 3 reviews rated 5 using NBP1-48308 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.1% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: PKM2

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.

Long Name

Pyruvate Kinase, Muscle

Alternate Names

CTHBP, OIP3, p58, PK2, PK3, TCB, THBP1, Tumor M2-PK

Entrez Gene IDs

5315 (Human); 18746 (Mouse)

Gene Symbol

PKM

UniProt

P14618 (Human), P52480 (Mouse)

Additional PKM2 Products

Product Documents for PKM2 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for PKM2 Antibody - BSA Free

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.

Citations for PKM2 Antibody - BSA Free

Customer Reviews for PKM2 Antibody - BSA Free (3)

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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • PKM2 Antibody
    Name: Sachin Puri
    Application: Immunocytochemistry
    Sample Tested: Human lung cancer cell lines
    Species: Human
    Verified Customer | Posted 06/15/2017
  • Name: Kelsey Stockbridge
    Application: Western Blot
    Sample Tested: HeLa cells
    Species: Human
    Verified Customer | Posted 02/17/2011
    PKM2 Antibody - BSA Free NBP1-48308
  • Name: Jian Zhang
    Application: Immunofluorescence
    Sample Tested: Rat glioblastoma frozen section
    Species: Rat
    Verified Customer | Posted 10/15/2010
    PKM2 Antibody - BSA Free NBP1-48308

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Protocols

View specific protocols for PKM2 Antibody - BSA Free (NBP1-48308):

Western Blot Protocol
1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% NFDM + 1% BSA in TBS + Tween, 1 hour at RT.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-PKM2 primary antibody (NBP1-48308) in blocking buffer and incubate 1 hour at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for PKM2 Antibody - BSA Free

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  • Q: I am wondering if this antibody can be used for IP or ChIP? Have you ever tested it in either of these applications?

    A:

    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.

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Associated Pathways

HIF Enhancer Pathways HIF Enhancer Pathway Thumbnail