PKM1 Antibody (PSH04-37)

Novus Biologicals | Catalog # NBP3-32825

Recombinant Monoclonal Antibody
Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # PSH04-37 expressed in HEK293
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Product Specifications

Immunogen

Synthetic peptide within human PKM1 aa 389-433 / 531 (P14618-2). (Uniprot: P14618-2)

Localization

Cytoplasm, Nucleus.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

58 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 PKM1 Antibody (PSH04-37)

PKM1 Antibody (PSH04-37)

Western Blot: PKM1 Antibody (PSH04-37) [NBP3-32825]

Western Blot: PKM1 Antibody (PSH04-37) [NBP3-32825] - Western blot analysis of PKM1 on different lysates with Rabbit anti-PKM1 antibody (NBP3-32825) at 1/1,000 dilution.

Lane 1: HCT 116 cell lysate (20 ug/Lane)
Lane 2: U-87 MG cell lysate (20 ug/Lane)
Lane 3: HeLa cell lysate (20 ug/Lane)
Lane 4: Human brain tissue lysate (40 ug/Lane)
Lane 5: Mouse brain tissue lysate (40 ug/Lane)
Lane 6: Mouse heart tissue lysate (40 ug/Lane)
Lane 7: Mouse liver tissue lysate (negative) (40 ug/Lane)

Predicted band size: 58 kDa
Observed band size: 55 kDa

Exposure time: 14 seconds; ECL;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32825) at 1/1,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
PKM1 Antibody (PSH04-37)

Immunohistochemistry: PKM1 Antibody (PSH04-37) [NBP3-32825]

Immunohistochemistry: PKM1 Antibody (PSH04-37) [NBP3-32825] - Immunohistochemical analysis of paraffin-embedded human heart tissue with Rabbit anti-PKM1 antibody (NBP3-32825) at 1/30,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32825) at 1/30,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
PKM1 Antibody (PSH04-37)

Immunocytochemistry/ Immunofluorescence: PKM1 Antibody (PSH04-37) [NBP3-32825]

Immunocytochemistry/ Immunofluorescence: PKM1 Antibody (PSH04-37) [NBP3-32825] - Immunocytochemistry analysis of HCT 116 cells labeling PKM1 with Rabbit anti-PKM1 antibody (NBP3-32825) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PKM1 antibody (NBP3-32825) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
PKM1 Antibody (PSH04-37)

Flow Cytometry: PKM1 Antibody (PSH04-37) [NBP3-32825]

Flow Cytometry: PKM1 Antibody (PSH04-37) [NBP3-32825] - Flow cytometric analysis of HCT 116 cells labeling PKM1.

Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32825, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Applications for PKM1 Antibody (PSH04-37)

Application
Recommended Usage

Flow Cytometry

1:1000

Immunocytochemistry/ Immunofluorescence

1:500

Immunohistochemistry-Paraffin

1:5000-1:30000

Western Blot

1:1000

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Advanced Features

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Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.1% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: PKM1

Pyruvate kinases are glycolytic enzymes that transfer a phosphate group from phosphoenolpyruvate to ADP generating pyruvate and ATP. There are 4 isozymes of pyruvate kinase in mammals: L, R, M1 and M2. L type is major isozyme in the liver, R is found in red cells, M1 is the main form in muscle, heart and brain, and M2 is found in early fetal tissues as well as in most cancer cells. The M1 isoform differs from the M2 isoform due to a change in sequence between residues 389-433. In the presence of D-fructose 1,6-bisphosphate (FBP) the monomer reversibly forms the highly active tetrameric form. This tetramer is associated with the glycolytic enzyme complex and activates the pyruvate kinase activity. A nearly inactive dimeric form is associates with tumor cells.

Alternate Names

PK1, Pyruvate kinase isozymes M1, Pyruvate kinase M1

Gene Symbol

PKM

Additional PKM1 Products

Product Documents for PKM1 Antibody (PSH04-37)

Certificate of Analysis

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Product Specific Notices for PKM1 Antibody (PSH04-37)

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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Protocols

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