PKM2 Antibody (YA6705)

Cat. No.: HY-P87012: Data Sheet COA
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PKM2 Antibody (YA6705) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to PKM2.

For research use only. We do not sell to patients.

Size	Stock	Quantity
10 μL	In-stock	Estimated Time of Arrival: December 31
50 μL	In-stock	Estimated Time of Arrival: December 31
100 μL	In-stock	Estimated Time of Arrival: December 31
250 μL	Get quote

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WB: Western Blot;
IHC-P: Immunohistochemistry-Paraffin;
IHC-F: Immunohistochemistry-Frozen;
ICC/IF: Immunocytochemistry/Immunofluorescence;
IF-Tissue: Immunofluorescence-Tissue;
mIHC: Multiplex Immunohistochemical;
IP: Immunoprecipitation;
ChIP: Chromatin Immunoprecipitation;
FC: Flow Cytometry;
ELISA: Enzyme Linked Immunosorbent Assay

Product Detail
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Background
Documentation

Description

PKM2 Antibody (YA6705) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to PKM2.

Host

Rabbit

Clonality

Recombinant,Monoclonal

Molecular Weight

Predicted band size: 58 kDa;

Observed band size: 58 kDa

Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.

Species Reactivity

Human, Mouse, Rat, Monkey

SwissProt ID

P14618

Gene ID

5315 [NCBI]

Immunogen

Synthetic peptide within human PKM2 aa 381-430.

Application &
Dilution Ratio

Application	Dilution Ratio
WB WB: Western Blot	1:10000
ICC/IF ICC/IF: Immunocytochemistry/Immunofluorescence	1:100
IHC-P IHC-P: Immunohistochemistry-Paraffin	1:1000
FC FC: Flow Cytometry	1:1000
IP IP: Immunoprecipitation	1-2μg/sample

Purity	affinity purified.	Conjugation	Non-conjugated
Modification	Unmodified	Isotype	IgG

Appearance

Liquid

Formulation

Supplied in PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Storage & Stability

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping

Shipping with blue ice.

Verification Image

ALL WB IHC-P mIHC FC

Western blot analysis of extracts fromHela (lane2(20μg), MCF-7 (lane3(20μg), A549 (lane4(20μg) and NIH3T3 (lane5(20μg) usingPKM2 Antibody (HY-P87012). Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/10000) and Loading control antibody (Beta Actin, HY-P80993, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human Colon cancer tissue using PKM2 antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P87012, 1:600 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Immunohistochemical analysis of paraffin-embedded human Prostate Cancer tissue using PKM2 antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P87012, 1:600 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Immunohistochemical analysis of paraffin-embedded human Endometrial Carcinoma tissue using PKM2 antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P87012, 1:600 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Immunohistochemical analysis of paraffin-embedded human Kidney cancer tissue using PKM2 antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P87012, 1:600 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Immunohistochemical analysis of paraffin-embedded human Ovarian Cancer‌ tissue using PKM2 antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P87012, 1:600 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Immunohistochemical analysis of paraffin-embedded human Gastric Cancer tissue using PKM2 antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P87012, 1:600 dilution) at room temperature for 60 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Colon cancer‌ tissue using PKM2 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P87012, 1:1000 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Colon cancer‌ tissue using PKM2 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P87012, 1:1000 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Colon cancer‌ tissue using PKM2 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P87012, 1:1000 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Liver Cancer tissue using PKM2 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P87012, 1:1000 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Liver Cancer tissue using PKM2 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P87012, 1:1000 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Liver Cancer tissue using PKM2 antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P87012, 1:1000 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.
Flow cytometric analysis of 1X10⁶ HeLa cells labeling PKM2 Antibody (HY-P87012, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/1000 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

Background

Function：Catalyzes the final rate-limiting step of glycolysis by mediating the transfer of a phosphoryl group from phosphoenolpyruvate (PEP) to ADP, generating ATP (PubMed:15996096, PubMed:1854723, PubMed:20847263). 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, PubMed:20847263). 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, PubMed:20847263)|Isoform specifically expressed during embryogenesis that has low pyruvate kinase activity by itself and requires allosteric activation by D-fructose 1,6-bisphosphate (FBP) for pyruvate kinase activity (PubMed:18337823, PubMed:20847263). In addition to its pyruvate kinase activity in the cytoplasm, also acts as a regulator of transcription in the nucleus by acting as a protein kinase (PubMed:18191611, PubMed:21620138, PubMed:22056988, PubMed:22306293, PubMed:22901803, PubMed:24120661). Translocates into the nucleus in response to various signals, such as EGF receptor activation, and homodimerizes, leading to its conversion into a protein threonine- and tyrosine-protein kinase (PubMed:22056988, PubMed:22306293, PubMed:22901803, PubMed:24120661, PubMed:26787900). Catalyzes phosphorylation of STAT3 at 'Tyr-705' and histone H3 at 'Thr-11' (H3T11ph), leading to activate transcription (PubMed:22306293, PubMed:22901803, PubMed:24120661). Its ability to activate transcription plays a role in cancer cells by promoting cell proliferation and promote tumorigenesis (PubMed:18337823, PubMed:22901803, PubMed:26787900). Promotes the expression of the immune checkpoint protein CD274 in BMAL1-deficient macrophages (By similarity). May also act 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 role in caspase independent cell death of tumor cells (PubMed:17308100)|Pyruvate kinase isoform expressed in adult tissues, which replaces isoform M2 after birth (PubMed:18337823). In contrast to isoform M2, has high pyruvate kinase activity by itself and does not require allosteric activation by D-fructose 1,6-bisphosphate (FBP) for activity (PubMed:20847263)

Subcellular Localization：Cytoplasm,Nucleus,Cytoplasm

Expression：
Tissue_Specificity: Specifically expressed in proliferating cells, such as embryonic stem cells, embryonic carcinoma cells, as well as cancer cells|Expressed in adult tissues (PubMed:18337823). Not expressed in tumor cells (PubMed:18337823)

Isoforms & Post-Translational Modification：P14618 has three isomers: P14618-1: 57937 Da (predicted); P14618-2: 58062 Da (predicted); P14618-3: 56273 Da (predicted).
ISGylated丨Under hypoxia, hydroxylated by EGLN3丨Acetylation at Lys-305 is stimulated by high glucose concentration, it decreases enzyme activity and promotes its lysosomal-dependent degradation via chaperone-mediated autophagy丨Acetylated at Lys-433 by EP300, leading to impair phosphoenolpyruvate substrate-binding and promote its homodimerization and subsequent translocation to the nucleus (PubMed:24120661)丨S-nitrosylation at Cys-423 and Cys-424 inhibits homotetramerization and pyruvate kinase activity (PubMed:30487609)丨FGFR1-dependent tyrosine phosphorylation is reduced by interaction with TRIM35

Subunit：Monomer and homotetramer; exists as a monomer in the absence of D-fructose 1,6-bisphosphate (FBP), and reversibly associates to form a homotetramer in the presence of FBP (PubMed:15996096, PubMed:18298799, PubMed:18337815, PubMed:1854723, PubMed:23064226, PubMed:2813362)

RRID

AB_3719247

Synonyms

CTHBP antibody; Cytosolic thyroid hormone-binding protein antibody; KPYM_HUMAN antibody; OIP-3 antibody; Opa-interacting protein 3 antibody; p58 antibody; pkm antibody; PKM1 antibody; PKM2 antibody; Pyruvate kinase 2/3 antibody; CTHBP antibody; Cytosolic thyroid hormone-binding protein antibody; KPYM_HUMAN antibody; OIP-3 antibody; Opa-interacting protein 3 antibody; p58 antibody; pkm antibody; PKM1 antibody; PKM2 antibody; Pyruvate kinase 2/3 antibody; Pyruvate kinase muscle isozyme antibody; Pyruvate kinase PKM antibody; THBP1 antibody; Thyroid hormone-binding protein 1 antibody; Tumor M2-PK antibody;

Documentation

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Data Sheet (231 KB) SDS (253 KB)

COA (206 KB)

User Guide for Antibodies (1077 KB)