Biolegend, 127401, Purified anti-mouse Podoplanin Antibody, 50μg

The mucin-type glycoprotein podoplanin is thought to be involved in the development of the lymphatic vascular system. Podoplanin is named after its expression in the kidney glomerular epithelial cells (podocytes). It has a potential role in tumor progression.
50μg
Verified Reactivity: Mouse
Antibody Type: Monoclonal
Host Species: Syrian Hamster
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.
Application: FC - Quality tested IHC-F - Verified SB - Reported in the literature, not verified in house
Recommended Usage: Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µl volume or 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes: Additional reported applications (for the relevant formats) include: immunohistochemistry6, and spatial biology (IBEX)8,9.
Additional Product Notes: Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
Application References(PubMed link indicates BioLegend citation): Farr A, et al. 1992. J. Histochem. Cytochem. 40:651. Farr AG, et al. 1992. J. Exp. Med. 176:1477. Bekiaris V, et al. 2008. J. Immunol. 180:6768. Algars A, et al. 2011. Blood 117:4387. PubMed Reis VO, et al. 2012. Immunobiology. 217:831. PubMed Kaji C, et al. 2012. Acta. Histochem. Cytochem. 45:227. (IHC) Kretschmer S, et al. 2013. PLoS One. 8:e52201. PubMed. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations: Kida H, et al. 2017. Sci Rep. 7:12032. PubMed Mahamud MR, et al. 2023. Microcirculation. 30:e12787. PubMed Lütge M, et al. 2023. Nat Immunol. . PubMed Hutton C, et al. 2021. Cancer Cell. 39:1227. PubMed Dubrot J, et al. 2018. Life Sci Alliance. 1:e201800164. PubMed Kretschmer S, et al. 2013. PLoS One. 8:e55201. PubMed Reis V, et al. 2012. Immunobiology. 217:831. PubMed Gao J, et al. 2017. Oncol Lett. 14:2954. PubMed Mahamud MR, et al. 2019. Development. 146:dev184218. PubMed Rodriguez AB, et al. 2021. Cell Reports. 36(3):109422. PubMed Kajiwara K, et al. 2021. BMC Nephrol. 22:3. PubMed Emgård J, et al. 2018. Immunity. 143:419. PubMed Davidson S, et al. 2020. Cell Reports. 31(7):107628. PubMed Hegde S, et al. 2020. Cancer Cell. 37(3):289-307. PubMed Cheng HW, et al. 2019. Nat Commun. 10:1739. PubMed Donati Y, et al. 2020. Am J Physiol Lung Cell Mol Physiol. L619:318. PubMed Molica F,et al. 2017. Sci Rep.. 10.1038/s41598-017-14130-4. PubMed Le HQ, et al. 2021. EMBO Rep. 22:e52785. PubMed ålgars A, et al. 2011. Blood. 117:4387. PubMed Chihara N, et al. 2018. Nature. 558:454. PubMed Munir H, et al. 2021. Nat Commun. 0.974305556. PubMed Alexandre YO, et al. 2020. Cell Reports. 33(13):108567. PubMed Kraft JC, et al. 2018. J Drug Target. 26:494. PubMed Kwok T, et al. 2022. Front Aging. 3:838943. PubMed Tamburini BAJ, et al. 2019. Front Immunol. 10:1036. PubMed Takeuchi Y, et al. 2021. Sci Rep. 11:18679. PubMed Blaskovic S, et al. 2020. Am J Physiol Lung Cell Mol Physiol. L606:318. PubMed Choi SY, et al. 2020. Nat Commun. 0.81875. PubMed Mece O, et al. 2022. Nat Commun. 13:2760. PubMed Nagashima K, et al. 2017. Biochem Biophys Res Commun. . 10.1016/j.bbrc.2017.09.004. PubMed
RRID: AB_1089186 (BioLegend Cat. No. 127401) AB_1089187 (BioLegend Cat. No. 127402)
Structure: 43 Kd glycosylated type-1 transmembrane protein. Mucin-type protein.
Distribution: Expressed on epithelial and mesothelial cells.
Cell Type: Epithelial cells
Biology Area: Cell Biology, Immunology, Neuroscience, Neuroscience Cell Markers
Antigen References: 1. Farr A, et al. 1992. J. Histochem. Cytochem. 40:651. 2. Schacht V, et al. 2005. Am. J. Pathol. 166:913.
Gene ID: 14726
UniProt: View information about Podoplanin on UniProt.org
Clone: 8.1.1
Regulatory Status: RUO
Other Names: T1a, gp38, stromal cell marker
Isotype: Syrian Hamster IgG
Q: If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?
A: It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.
Q: Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?
A: Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.
Q: Are other fluorophores compatible with IBEX?
A: Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.
Q: The same antibody works in one tissue type but not another. What is happening?
A: Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.
Q: How can I be sure the staining I’m seeing in my tissue is real?
A: In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.