Human/Rat PKM1/2 Antibody

Name: Human/Rat PKM1/2 Antibody
Brand: R&D Systems
SKU: MAB7244

R&D Systems, part of Bio-Techne | Catalog # MAB7244

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human, Rat

Applications

Immunocytochemistry, Immunohistochemistry, Knockout Validated, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG_2B Clone # 937801

View all PKM2 Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human PKM2
Ser2-Pro531
Accession # P14618-1

Specificity

Detects human PKM2 in direct ELISAs. Detects human and rat PKM1/2 in Western blot.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2B

Scientific Data Images for Human/Rat PKM1/2 Antibody

PKM2 in WI‑38 Human Cell Line.

PKM2 was detected in immersion fixed WI-38 human lung fibroblast cell line using Mouse Anti-Human/Rat PKM1/2 Monoclonal Antibody (Catalog # MAB7244) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

PKM2 in Human Prostate.

PKM2 was detected in immersion fixed paraffin-embedded sections of human prostate using Mouse Anti-Human/Rat PKM1/2 Monoclonal Antibody (Catalog # MAB7244) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to epithelial cell cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

PKM2 in Rat Skeletal Muscle.

PKM2 was detected in perfusion fixed frozen sections of rat skeletal muscle using Mouse Anti-Human/Rat PKM1/2 Monoclonal Antibody (Catalog # MAB7244) at 8 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to epithelial cell cytoplasm. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Detection of Human PKM2 by Simple Western^TM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for PKM2 at approximately 61 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Rat PKM1/2 Monoclonal Antibody (Catalog # MAB7244). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western Blot Shows Human PKM2 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and PKM2 knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Rat PKM1/2 Monoclonal Antibody (Catalog # MAB7244) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for PKM2 at approximately 60 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (AF5718) is shown as a loading control.. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

PKM2 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

PKM2 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line but is not detected in PKM2 knockout (KO) HeLa Human Cell Line cell line using Mouse Anti-Human/Rat PKM1/2 Monoclonal Antibody (Catalog # MAB7244) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; NL009) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human PKM2 by Western Blot.

Western blot shows lysates of U-87 MG human glioblastoma/astrocytoma cell line, A549 human lung carcinoma cell line, HeLa human cervical epithelial carcinoma cell line, human skeletal muscle. PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human/Rat PKM2 Monoclonal Antibody (Catalog # MAB7244) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for PKM2 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

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Applications for Human/Rat PKM1/2 Antibody

Application

Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed WI‑38 human lung fibroblast cell line

Immunohistochemistry

1-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human prostate and perfusion fixed frozen sections of rat skeletal muscle

Knockout Validated

PKM2 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in PKM2 knockout HeLa cell line.

Simple Western

10 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line

Western Blot

1 µg/mL
Sample: A549 human lung carcinoma cell line, HeLa human cervical epithelial carcinoma cell line, U‑87 MG human glioblastoma/astrocytoma cell line, human skeletal muscle

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: PKM2

PKM2 (Pyruvate Kinase isoenzyme M2; also p58, OIP3, THBP1 and CTHBP) is a 58-60 kDa member of the PK family of enzymes. It is widely expressed, being found both intracellularly and in blood, and represents the more common splice variant of the PKM gene. PKM2 generates ATP and pyruvate by catalyzing the transfer of a phosphoryl group from PEP to ADP. Thus, when active, PKM2 promotes energy production and glycolysis. PKM2 exists as a marginally active monomer, with full activity achieved through homotetramerization. Notably, in tumor cells, select oncogenes appear to induce PKM2 homodimerization which limits PKM2 activity. PKM2 is known to be regulated by the binding of T3 and Fru-1,6-bisP. Human PKM2 is 531 amino acids (aa) in length. It contains a catalytic region (aa 43‑527) plus four utilized Ser/Thr and Tyr phosphorylation sites, respectively. PKM1 is another PKM gene splice variant that shows a 45 aa substitution for
aa 389‑433 of PKM2. This variant shows limited expression (striated muscle) and hyperbolic Michaelis-Menten kinetics. There are additional isoform variants of PKM2 that show either a deletion of aa 59‑132, or a 67 aa substitution for aa 1‑82. Over aa 434‑531, human PKM2 shares 95% aa sequence identity with mouse PKM2.

Long Name

Pyruvate Kinase, Muscle

Alternate Names

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

Entrez Gene IDs

5315 (Human); 18746 (Mouse); 25630 (Rat)

Gene Symbol

PKM

UniProt

P14618-1

Additional PKM2 Products

Product Documents for Human/Rat PKM1/2 Antibody

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COA

SDS

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Product Specific Notices for Human/Rat PKM1/2 Antibody

For research use only

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