AB214257

Alexa Fluor® 647 Anti-PKM antibody [EPR10138(B)]

Name: Anti-PKM Alexa Fluor® 647 antibody [EPR10138(B)] (ab214257) IgG | Abcam
Brand: Abcam Limited
SKU: AB214257
Price: 185 USD
Availability: InStock
Rating: 4 (1 reviews)

RabMAb
Recombinant
What is this?

(1 Review)

(1 Publication)

Rabbit Recombinant Monoclonal KPYM antibody - conjugated to Alexa Fluor® 647. Suitable for ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

View Alternative Names

OIP3, PK2, PK3, PKM2, PKM, Pyruvate kinase PKM, Cytosolic thyroid hormone-binding protein, Opa-interacting protein 3, Pyruvate kinase 2/3, Pyruvate kinase muscle isozyme, Threonine-protein kinase PKM2, Thyroid hormone-binding protein 1, Tumor M2-PK, Tyrosine-protein kinase PKM2, p58, CTHBP, OIP-3, THBP1

2 Images

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-PKM antibody [EPR10138(B)] (AB214257)

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-PKM antibody [EPR10138(B)] (AB214257)

Flow cytometry scatter plots showing human peripheral mononuclear cells (PBMCs), either stained with ab214257 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab214257) (1x 10⁶ in 100μl at 0.2 μg/ml (1/2500)) for 30min on ice. Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-PKM antibody [EPR10138(B)] (AB214257)

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-PKM antibody [EPR10138(B)] (AB214257)

ab214257 staining PKM in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), and then incubated in 1%BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated overnight at +4°C with ab214257 at 1/50 dilution (shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).

Unconjugated

Anti-PKM antibody [EPR10138(B)]
Carrier free

Anti-PKM antibody [EPR10138(B)] - BSA and Azide free
660 APC

APC Anti-PKM antibody [EPR10138(B)]
565 Alexa Fluor® 555

Alexa Fluor® 555 Anti-PKM antibody [EPR10138(B)]
617 Alexa Fluor® 594

Alexa Fluor® 594 Anti-PKM antibody [EPR10138(B)]
578 PE

PE Anti-PKM antibody [EPR10138(B)]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR10138(B)

Isotype

IgG

Conjugation

Alexa Fluor® 647

Excitation/Emission

Ex: 650nm, Em: 665nm

Carrier free

Reacts with

Human, Human, Mouse, Rat

Applications

ICC/IF, Flow Cyt (Intra)

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/100", "ICCIF-species-notes": "This product gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min)", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Mouse": { "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Rat": { "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Pig": { "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" } } }

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Product details

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.4 Preservative: 0.02% Sodium azide Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle|Store in the dark

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

PKM also known as pyruvate kinase muscle isozyme (PKM) and PEP is an enzyme that plays an important role in glycolysis by catalysing the conversion of phosphoenolpyruvate (PEP) to pyruvate yielding ATP in the process. The PKM protein has two isoforms PKM1 and PKM2 which result from alternative splicing of the PKM gene. The mass of PKM2 the more studied isoform is approximately 58 kDa. PKM is expressed in various tissues prominently in skeletal muscle heart brain and many tumor cells. Additionally PKM has significant activity in rapidly proliferating cells suggesting its importance in high-energy demanding environments.

Biological function summary

PKM functions not only in catalyzing the last step of glycolysis but also regulates metabolic and transcriptional processes. Specifically PKM2 is a participant in the regulation of gene expression and cellular response to oxidative stress and nutrient availability. It can exist as a dimer or tetramer with the latter being the more active form in glycolytic pathways while the dimeric form can translocate to the nucleus to perform functions unrelated to its glycolytic activity. These transformations make PKM part of a dynamic complex that responds to various cellular signals.

Pathways

PKM integrates into essential metabolic pathways including the glycolytic pathway and influences the pentose phosphate pathway. It works in conjunction with phosphofructokinase-1 (PFK1) another key glycolytic enzyme synchronizing the energy production process in cells. PKM2's non-metabolic roles involve interactions in signaling pathways related to cellular proliferation and survival often interacting with and modulating proteins like HIF-1α which plays a central role in cellular responses to hypoxia.

PKM2 shows strong connections to cancer and metabolic diseases. Tumor cells often exhibit a shift in expression from PKM1 to PKM2 facilitating the altered metabolism known as the Warburg effect characterized by increased aerobic glycolysis. Its interaction with HIF-1α promotes adaptation to low oxygen environments typical in tumorous growth. Furthermore PKM disruptions or aberrations contribute to metabolic disorders such as diabetes where altered glucose metabolism becomes evident. The protein's behavior in these disease conditions indicates potential targets for therapeutic intervention highlighting the importance of PKM in both normal physiology and pathology.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

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 M2. 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).. Isoform M1. 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).

See full target information PKM

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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Cell metabolism 29:856-870.e7 PubMed30686744

2019

Polyunsaturated Fatty Acid Desaturation Is a Mechanism for Glycolytic NAD Recycling.

Applications

Unspecified application

Species

Unspecified reactive species

Wondong Kim,Amy Deik,Clicerio Gonzalez,Maria Elena Gonzalez,Feifei Fu,Michele Ferrari,Claire L Churchhouse,Jose C Florez,Suzanne B R Jacobs,Clary B Clish,Eugene P Rhee

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Alexa Fluor® 647 Anti-PKM antibody [EPR10138(B)]

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-PKM antibody [EPR10138(B)] (AB214257)

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-PKM antibody [EPR10138(B)] (AB214257)

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Key facts

Host species

Clonality

Clone number

Isotype

Conjugation

Excitation/Emission

Carrier free

Reacts with

Applications

Immunogen

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Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (1)

Product promise