PKM2-specific Recombinant antibody

PKM2-specific Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

241431E8

Cat no : 84451-1-RR

Synonyms

PKM, PKM1, PK-M1, PKM1 pyruvate kinase, muscle, PKM2

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Validation Data Gallery

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  WB analysis using 84451-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 84451-1-RR (PK-M2-specific antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IHC staining of human colon cancer using 84451-1-RR

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 84451-1-RR (PKM2-specific antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon cancer using 84451-1-RR

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 84451-1-RR (PKM2-specific antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 84451-1-RR

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using PKM2-specific antibody (84451-1-RR, Clone: 241431E8 ) at dilution of 1:250 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of HeLa using 84451-1-RR

  1x10^6 HeLa cells were intracellularly stained with 0.25 ug PKM2-specific Recombinant antibody (84451-1-RR, Clone:241431E8) and APC-Conjugated Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  FC experiment of HepG2 using 84451-1-RR

  1x10^6 HepG2 cells were intracellularly stained with 0.25 ug PKM2-specific Recombinant antibody (84451-1-RR, Clone:241431E8) and APC-Conjugated Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: U2OS cells, HeLa cells, HEK-293 cells, A549 cells, MDA-MB-231 cells
• Positive IHC detected in: human colon cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: HepG2 cells, HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:125-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

84451-1-RR targets PKM2-specific in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Peptide
• Full Name: PKM pyruvate kinase, muscle
• Calculated Molecular Weight: 58 kDa
• Observed Molecular Weight: 58 kDa
• GenBank Accession Number: NM_002654
• Gene Symbol: PKM
• Gene ID (NCBI): 5315
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

PKM, also named as OIP3, PK2, PK3, PKM, p58, THBP1, CTHBP and Tumor M2-PK, belongs to the pyruvate kinase family. It is glycolytic enzyme that catalyzes the transfer of a phosphoryl group from phosphoenolpyruvate (PEP) to ADP, generating ATP. It stimulates POU5F1-mediated transcriptional activation. PKM plays a general role in caspase independent cell death of tumor cells. PKM has 2 isoforms named as PKM1/M2. The activity of the M2 isoform (but not the M1 isoform) can be inhibited by tyrosine kinase signalling in tumourcells. The primary pyruvate kinase isoform before tumour development is PK-M1; however, the primary isoform from four independent tumours is PK-M2 (PMID:18337823). The immunogen of this antibody is M2 isoform and this antibody is specific to PKM2 isoform.