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Pyruvate Kinase M1/2 Peptide 1

Antibody: CPTC-PKM-1
Background

Catalog Number:

CPTC-PKM-1

Target Antigen:

Pyruvate Kinase M1/2 Peptide 1

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

02/09/2024

Antigen Recognition(s):

Peptide, Recombinant Full-length, Endogenous, Phosphorylation

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Characterization Data [Compare Characterization Data]
  Affinity Measurement by Biolayer Interferometry
Click to enlarge image The affinity and binding kinetics of CPTC-PKM-1 and BSA-conjugated peptide “RLDID-pS-PPIT" were measured using biolayer interferometry. CPTC-PKM-1 antibody was captured onto Protein A biosensors. BSA-conjugated peptide, 0.25 nM, 0.0635 nM, 0156 nM, and 0.0039 nM, was used as analyte. Buffer only and biosensors immobilized without antibody were used as references for background subtraction. Click image to enlarge

CPTC-PKM-1 Affinity and Kinetics (BLI)

Result: Positive

The affinity and binding kinetics of CPTC-PKM-1 and BSA-conjugated peptide “RLDID-pS-PPIT" were measured using biolayer interferometry. CPTC-PKM-1 antibody was captured onto Protein A biosensors. BSA-conjugated peptide, 0.25 nM, 0.0635 nM, 0156 nM, and 0.0039 nM, was used as analyte. Buffer only and biosensors immobilized without antibody were used as references for background subtraction.


Characterization SOP Files

  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-PKM-1 antibody and BSA-conjugated peptide, "RLDID-pS-PPIT", were measured using surface plasmon resonance. BSA-conjugated peptide was amine coupled on a Series S CM5 biosensor chip. PKM-1 antibody was titrated using 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM, and 0.0625 nM as analyte. Binding data were double-referenced and analyzed globally.  Kinetic constants were difficult to determine.   Click image to enlarge

CPTC-PKM-1 Affinity and Kinetics (Surface Plasmon Resonance)  

Result: Positive

Affinity and binding kinetics of CPTC-PKM-1 antibody and BSA-conjugated peptide, "RLDID-pS-PPIT", were measured using surface plasmon resonance. BSA-conjugated peptide was amine coupled on a Series S CM5 biosensor chip. PKM-1 antibody was titrated using 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM, and 0.0625 nM as analyte. Binding data were double-referenced and analyzed globally.  Kinetic constants were difficult to determine.

 


Characterization SOP Files

  IHC Tissue
Click to enlarge image Tissue Micro-Array (TMA) core of colon cancer showing nuclear and cytoplasmic staining using Antibody CPTC-PKM-1. Titer: 1:200 Click image to enlarge

CPTC-PKM-1 IHC Tissue

Result: Positive

Tissue Micro-Array (TMA) core of colon cancer showing nuclear and cytoplasmic staining using Antibody CPTC-PKM-1. Titer: 1:200


Characterization SOP Files

  Immunofluorescence
Click to enlarge image Immunofluorescence staining of human cell line OVCAR-3 using CPTC-PKM-1 as primary antibody. PKM protein expression shows localization in the nucleus and cytoplasm (green). Click image to enlarge

CPTC-PKM-1 Immunofluorescence

Result: Presumed Positive

Immunofluorescence staining of human cell line OVCAR-3 using CPTC-PKM-1 as primary antibody. PKM protein expression shows localization in the nucleus and cytoplasm (green).


Characterization SOP Files

  Immunofluorescence
Click to enlarge image Immunofluorescence staining of human cell line OVCAR-3 using CPTC-PKM-1 as primary antibody. PKM protein expression shows localization in the nucleus and cytoplasm (green). Click image to enlarge

CPTC-PKM-1 Immunofluorescence

Result: Presumed Positive

Immunofluorescence staining of human cell line OVCAR-3 using CPTC-PKM-1 as primary antibody. PKM protein expression shows localization in the nucleus and cytoplasm (green).


Characterization SOP Files

  Indirect ELISA
Click to enlarge image Indirect ELISA using CPTC-PKM-1 antibody against BSA-conjugated peptide, "RLDID-pS-PPIT".  Click image to enlarge

CPTC-PKM-1 Indirect ELISA

Result: Positive

Indirect ELISA using CPTC-PKM-1 antibody against BSA-conjugated peptide, "RLDID-pS-PPIT". 


Characterization SOP Files

  NCI 60 Protein Array

CPTC-PKM-1 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


Characterization SOP Files

  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Automated western blot using CPTC-PKM-1 as primary antibody against recombinant not-phosphorylated PKM (PKM2 WT) and phosphorylated PKM (PKM2 pS37). The antibody was able to specifically recognize the phosphorylated recombinant protein. Click image to enlarge

CPTC-PKM-1 Automated WB (recombinant protein)

Result: Positive

Automated western blot using CPTC-PKM-1 as primary antibody against recombinant not-phosphorylated PKM (PKM2 WT) and phosphorylated PKM (PKM2 pS37). The antibody was able to specifically recognize the phosphorylated recombinant protein.


Characterization SOP Files

  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Western blot using CPTC-PKM-1 as primary antibody against recombinant not-phosphorylated PKM (PKM2 WT) and hosphorylated PKM (PKM2 pS37). The antibody was able to specifically recognize the phosphorylated recombinant protein. Weak signal. Click image to enlarge

CPTC-PKM-1 Western Blot (recombinant protein)

Result: Positive

Western blot using CPTC-PKM-1 as primary antibody against recombinant not-phosphorylated PKM (PKM2 WT) and hosphorylated PKM (PKM2 pS37). The antibody was able to specifically recognize the phosphorylated recombinant protein. Weak signal.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate

CPTC-PKM-1 Single Cell Western Blot (Cell Lysate)

Result: Negative

Single cell western blot using CPTC-PKM-1 as a primary antibody against OVCAR-3 cell lysate. PKM protein expression was not detected.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated western blot using CPTC-PKM-1 as primary antibody against Whole lysate of OVCAR3 cells. The antibody was able to presumebly detect the target protein in the tested lysate. Click image to enlarge

CPTC-PKM-1 Automated WB (lysate)

Result: Presumed Positive (with additional bands)

Automated western blot using CPTC-PKM-1 as primary antibody against Whole lysate of OVCAR3 cells. The antibody was able to presumebly detect the target protein in the tested lysate.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated Western Blot (Simple Western) using CPTC-PKM-1 as primary antibody against a selection of human lung cancer tissues, for which proteomics data have already shown PKM expression at different level. The antibody was able to detect target protein expression in all the lung tissue samples at different level. Proteomics data can be found at https://pdc.cancer.gov/pdc/study/PDC000149 Click image to enlarge

CPTC-PKM-1 Automated Western Blot (Tissue Lysates)

Result: Positive

Automated Western Blot (Simple Western) using CPTC-PKM-1 as primary antibody against a selection of human lung cancer tissues, for which proteomics data have already shown PKM expression at different level. The antibody was able to detect target protein expression in all the lung tissue samples at different level.
Proteomics data can be found at https://pdc.cancer.gov/pdc/study/PDC000149


Characterization SOP Files

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Antigen: Pyruvate Kinase M1/2 Peptide 1
Background

NCI Identification Number:

00516

Antigen Name:

Pyruvate Kinase M1/2 Peptide 1

CPTC Name:

CPTC-PKM Peptide 1

Aliases:

Pyruvate Kinase M1/2; THBP1; OIP3; PK3; PKM2; Cytosolic Thyroid Hormone-Binding Protein; Thyroid Hormone-Binding Protein 1; Pyruvate Kinase Muscle Isozyme; Pyruvate Kinase, Muscle; Pyruvate Kinase PKM; Pyruvate Kinase 2/3; Tumor M2-PK; EC 2.7.1.40; CTHBP; P58; Thyroid Hormone-Binding Protein, Cytosolic; Epididymis Secretory Protein Li 30; Pyruvate Kinase Isozymes M1/M2; OPA-Interacting Protein 3; Opa-Interacting Protein 3; PK, Muscle Type; HEL-S-30; OIP-3; TCB; PK2

Function:

This gene encodes a protein involved in glycolysis. The encoded protein is a pyruvate kinase that catalyzes the transfer of a phosphoryl group from phosphoenolpyruvate to ADP, generating ATP and pyruvate. This protein has been shown to interact with thyroid hormone and may mediate cellular metabolic effects induced by thyroid hormones. This protein has been found to bind Opa protein, a bacterial outer membrane protein involved in gonococcal adherence to and invasion of human cells, suggesting a role of this protein in bacterial pathogenesis. Several alternatively spliced transcript variants encoding a few distinct isoforms have been reported.PKM (Pyruvate Kinase M1/2) is a Protein Coding gene. Diseases associated with PKM include Pyruvate Kinase Deficiency Of Red Cells and Anterior Dislocation Of Lens. Among its related pathways are Development Beta-adrenergic receptors regulation of ERK and Metabolism. Gene Ontology (GO) annotations related to this gene include RNA binding and MHC class II protein complex binding. An important paralog of this gene is PKLR.Glycolytic enzyme that catalyzes the transfer of a phosphoryl group from phosphoenolpyruvate (PEP) to ADP, generating ATP (PubMed:15996096, PubMed:1854723). The ratio between the highly active tetrameric form and nearly inactive dimeric form determines whether glucose carbons are channeled to biosynthetic processes or used for glycolytic ATP production (PubMed:15996096, PubMed:1854723). The transition between the 2 forms contributes to the control of glycolysis and is important for tumor cell proliferation and survival (PubMed:15996096, PubMed:1854723). In addition to its role in glycolysis, also regulates transcription (PubMed:18191611, PubMed:21620138). Stimulates POU5F1-mediated transcriptional activation (PubMed:18191611). Promotes in a STAT1-dependent manner, the expression of the immune checkpoint protein CD274 in ARNTL/BMAL1-deficient macrophages (By similarity). Also acts as a translation regulator for a subset of mRNAs, independently of its pyruvate kinase activity: associates with subpools of endoplasmic reticulum-associated ribosomes, binds directly to the mRNAs translated at the endoplasmic reticulum and promotes translation of these endoplasmic reticulum-destined mRNAs (By similarity). Plays a general role in caspase independent cell death of tumor cells.

Chromosomal Localization:

15q23

Accession Number:

NP_001193725.1

UniProt Accession Number:

P14618

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

RLDID(pS)PPIT

Native Sequence:

Calculated Isoelectric Point:

Molecular Weight:

1210

Last Updated:

05/31/2022

Links

Characterization Data

SOPs

No SOPs available.

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