PE Rat Anti-Human Podoplanin (PA Tag)

BD Pharmingen™ PE Rat Anti-Human Podoplanin (PA Tag)

Name: PE Rat Anti-Human Podoplanin (PA Tag)
Brand: BD Pharmingen™
SKU: 571913

Clone NZ-1

(RUO)

Flow cytometric analysis of PA Tag expression in transfected cells. Transfected cells with an intracellular protein linked with PA were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with Perm/Wash Buffer (Cat. No. 554723). The cells were then stained with either BD Pharmingen™ PE Rat IgG2a, λ Isotype Control (Cat. No. 557076; dashed lined histogram) or BD Pharmingen™ PE Rat Anti-Human Podoplanin (PA Tag) antibody (Cat. No. 571912/571913; solid line histogram) at 0.25 µg/test. The fluorescence histogram showing PA Tag expression (or Ig Isotype staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis was 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 Podoplanin (PA Tag) expression on viable NCCIT cells. Cells from the Human NCCIT (Pluripotent embryonal carcinoma, ATCC® CRL-2073™) cell line were stained with either BD Pharmingen™ PE Rat IgG2a, λ Isotype Control (Cat. No. 557076; dashed lined histogram) or BD Pharmingen™ PE Rat Anti-Human Podoplanin (PA Tag) antibody (Cat. No. 571912/571913; solid line histogram) at 0.25 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing Podoplanin (PA Tag) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.

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Product Details

Brand: BD Pharmingen™

Alternative Name: AGGRUS; GP36; GP40; Gp38; PA2.26 antigen; PDPN; T1A

Reactivity: Human (QC Testing)

Isotype: Rat IgG2a, λ

Immunogen: Human Podoplanin Peptide (aa 38-51)

Application: Flow cytometry (Routinely Tested), Intracellular staining (flow cytometry) (Tested During Development)

Concentration: 0.2 mg/ml

Entrez Gene ID: 10630

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: 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 unconjugated antibody and free dye were 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.
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.
This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
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.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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

PE Rat IgG2a, λ Isotype Control RUO

Size 0.1 mg Cat No. 557076

Fixation Buffer RUO

Size 100 mL Cat No. 554655

Perm/Wash Buffer RUO

Size 100 mL Cat No. 554723

571913 Rev. 1

Antibody Details

NZ-1

The NZ-1 monoclonal antibody specifically binds with high affinity to Podoplanin. This antibody was generated by immunizing mice with a 14-residue peptide segment (aa 38-51) from a platelet aggregation-stimulating (PLAG) domain of human Podoplanin. For this reason, this antibody is useful to detect the Podoplanin molecule as well as target proteins, such as antibodies that are labeled with a dodecapeptide (GVAMPGAEDDVV) named PA Tag. Podoplanin is a ~43 kDa mucin-type 1 transmembrane glycoprotein composed of a serine- and threonine-rich intracellular domain, a single transmembrane domain and a short intracellular domain. This multifunctional glycoprotein is expressed on various cell types, including intestinal and thymic epithelial cells, lymphatic endothelial cells, kidney podocytes, alveolar type I cells, fibroblastic reticular cells, macrophages, Th17 cells and osteocytes. Podoplanin expression is upregulated in inflammation and cancer. The binding of Podoplanin to the C-type lectin domain family 1 member B (Clec1B, also known as CLEC-2) on the surface of platelets results in platelet aggregation/activation, thrombosis, lymphatic vessel development, and cancer invasion and metastasis. The Podoplanin (PA Tag) antibody can be used for detecting tagged target proteins in cells that do not express endogenous Podoplanin. This antibody specifically reacts with the human Podoplanin sequence and does not react with Podoplanin sequences from mouse, rat, hamster or dog. Reportedly, this antibody can be used for Western blot, immunohistochemistry, immunoprecipitation or imaging applications.

571913 Rev. 1

Format Details

R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE

Excitation Source: Yellow-Green 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

571913 Rev.1

Citations & References

View product citations for antibody "571913" on CiteAb

Development References (5)

Brown ZP, Takagi J. The PA Tag: A Versatile Peptide Tagging System in the Era of Integrative Structural Biology.. Adv Exp Med Biol. 2018; 1105:59-76. (Clone-specific: Western blot). View Reference
Fujii Y, Kaneko M, Neyazaki M, Nogi T, Kato Y, Takagi J. PA tag: a versatile protein tagging system using a super high affinity antibody against a dodecapeptide derived from human podoplanin.. Protein Expr Purif. 2014; 95:240-7. (Clone-specific: Flow cytometry, Western blot). View Reference
Fujii Y, Matsunaga Y, Arimori T, et al. Tailored placement of a turn-forming PA tag into the structured domain of a protein to probe its conformational state.. J Cell Sci. 2016; 129(7):1512-22. (Clone-specific: Immunoprecipitation). View Reference
Kato Y, Kaneko MK, Kuno A, et al. Inhibition of tumor cell-induced platelet aggregation using a novel anti-podoplanin antibody reacting with its platelet-aggregation-stimulating domain.. Biochem Biophys Res Commun. 2006; 349(4):1301-7. (Immunogen: Flow cytometry, Immunohistochemistry, Western blot). View Reference
Tamura-Sakaguchi R, Aruga R, Hirose M, et al. Moving toward generalizable NZ-1 labeling for 3D structure determination with optimized epitope-tag insertion.. Acta Crystallogr D Struct Biol. 2021; 77(Pt 5):645-662. (Clone-specific: Immunoprecipitation). View Reference

View All (5)

571913 Rev. 1

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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.