Flow cytometric analysis of PKM expression in HEK293T cells. Cells from the Human HEK293T (Embryonic Kidney, ATCC® CRL-3216 ™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then stained with Alexa Fluor™ 647 Rabbit IgG Isotype Control (Cat. No. 569345; dashed line histogram) or Alexa Fluor™ 647 Rabbit Anti-Human PKM antibody (Cat. No. 570749; solid line histogram) at 0.125 µg/test. The fluorescence histogram showing PKM expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of PKM expression in HEK293T cells. Cells from the Human HEK293T (Embryonic Kidney, ATCC® CRL-3216 ™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then stained with Alexa Fluor™ 647 Rabbit IgG Isotype Control (Cat. No. 569345; dashed line histogram) or Alexa Fluor™ 647 Rabbit Anti-Human PKM antibody (Cat. No. 570749; solid line histogram) at 0.125 µg/test. The fluorescence histogram showing PKM expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
CTHBP; OIP3; OPA-interacting protein 3; PK, muscle type; PKM; PKM2
Human (QC Testing), Mouse,Rat (Reported)
Rabbit IgG
Human PKM (aa 50-150)
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
5315
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- This rabbit monoclonal (RabMabTM) product is, in part, provided under an intellectual property license from Abcam. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product for buyer’s internal research use only (whether the buyer is an academic or for-profit entity) and buyer shall not be permitted to reverse engineer or otherwise perform any compositional, structural, functional or other analyses directed to learning the methodology, formulae, sequences, process, make-up or production of this product (or any portion thereof). 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: (i) 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. If you, as the buyer, do not agree to these conditions, please leave the product unopened and return the contents of this package to BD Biosciences, 10975 Torreyana Road, San Diego, CA 92121. For information on purchasing a license to this product for purposes other than research, contact Abcam at partnerships@abcam.com.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Alexa Fluor™ 647 Rabbit IgG Isotype Control RUO
Size
25 µg
Cat No.
569345
570749 Rev. 1
Antibody Details
EPR10138 (B)
The EPR10138 (B) recombinant monoclonal antibody specifically recognizes Pyruvate kinase (PKM), which is also known as Cytosolic thyroid hormone-binding protein (CTHBP) or OPA-interacting protein 3 (OIP3). PKM is a cytosolic enzyme that catalyzes the irreversible conversion of phosphoenolpyruvate and ADP to ATP and pyruvic acid in the last step of glycolysis. PKM has two isoforms, PKM1 and PKM2, which represent two splice variants of PKM. PKM1 is found in tissues with high catabolic demand, including non-proliferative tissues and muscle; the embryonic isoform, PKM2, is expressed in cancer, where it plays a central role in caspase-independent cell death of tumor cells.
570749 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
570749 Rev.1
Citations & References
View product citations for antibody "570749" on CiteAb
Development References (4)
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Fan XG, Pei SY, Zhou D, et al. Melittin ameliorates inflammation in mouse acute liver failure via inhibition of PKM2-mediated Warburg effect.. Acta Pharmacol Sin. 2021; 42(8):1256-1266. (Clone-specific: Immunofluorescence, Western blot). View Reference
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Israelsen WJ, Vander Heiden MG. Pyruvate kinase: Function, regulation and role in cancer.. Semin Cell Dev Biol. 2015; 43:43-51. (Biology). View Reference
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Steták A, Veress R, Ovádi J, Csermely P, Kéri G, Ullrich A. Nuclear translocation of the tumor marker pyruvate kinase M2 induces programmed cell death.. Cancer Res. 2007; 67(4):1602-8. (Biology). View Reference
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Wang C, He C, Lu S, et al. Autophagy activated by silibinin contributes to glioma cell death via induction of oxidative stress-mediated BNIP3-dependent nuclear translocation of AIF.. Cell Death Dis. 2020; 11(8):630. (Clone-specific: Western blot). View Reference
570749 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.