PODXL Mouse Monoclonal Antibody [Clone ID: 53D11]

Specifications

Product Data
Clone Name	53D11
Applications	FC, IHC
Recommended Dilution	Flow cytometry: 10-20 µg/ml (final concentration). For details see protocol below. Immunohistochemistry: Clone 53D11 has been reported by customer to stain Podocalyxin in Formalin-Fixed, Paraffin-Embedded human kidney tissue at an antibody concentration of 10 µg/ml.
Reactivities	Human
Host	Mouse
Isotype	IgG2a
Clonality	Monoclonal
Immunogen	CHO cell expressing full length human Podocalyxin/PCLP1
Specificity	This antibody reacts with human Podocalyxin/PCLP1.
Formulation	PBS containing 50% glycerol, pH 7.2. No preservative is contained. State: Azide Free State: Liquid Ig fraction
Concentration	lot specific
Purification	Protein A agarose
Conjugation	Unconjugated
Storage	Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing.
Stability	Shelf life: One year from despatch.
Database Link	Entrez Gene 5420 Human UniProt ID O00592
Background	Recent studies with avian embryos and murine embryonic stem cells have suggested that hematopoietic cells are derived from hemangioblasts, the common precursors of hematopoietic and endothelial cells. Hara et al. molecularly clon ed podocalyxin-like protein 1 (PCLP1) as a novel surface marker for endothelial-like cells in the AGM (aorta-gonad-mesonephros) region of mouse embryos, where long-term repopulating hematopoietic stem cells (LTR-HSCs) are known to arise. PCLP1 + CD45 - cells in the AGM region incorporated acetylated low-density lipopro tein and produced both hematopoietic and endothelial cells when cocultured with OP9 stromal cells. Moreover, multiple lineages of hematopoietic cells were generated in vivo when PCLP1 + CD45 - cells were injected into neonatal liver of busulfan-treated mice. Today it is reported that the PCLP1 is identical with the Podocalyxin.
Synonyms	Podocalyxin-like protein 1, PCLP, PCLP1, PCLP-1, GCTM-2 antigen, Hematopoietic Stem Cell Marker
Note	This product was originally produced by MBL International. Protocol: Flow cytometric analysis for floating cells We usually use Fisher tubes or equivalents as reaction tubes for all step described below. 1) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS) and 0.1% NaN 3]. 2) Resuspend the cells with washing buffer (5x10e6 cells/mL). 3) Add 50 μ L of the cell suspension into each tube, and centrifuge at 500 x g for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 4) Add 10 μ L of Clear Back (human Fc receptor blocking reagent) and 0.1% NaN 3 to the cell pellet after tapping. Mix well and incubate for 5 minutes at room temperature. 5) Add 30 μL of the Anti-Human Podocalyxin/PCLP1 monoclonal antibody (53D11) (10-20 μ g/mL) diluted with the washing buffer. Mix well and incubate for 30 minutes at room temperature. 6) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration. 7) Add 30 μL of secondary antibody (1:40 FITC conjugated anti-mouse IgG) diluted with the washing buffer. Mix well and incubate for 15 minutes at room temperature. 8) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration. 9) Resuspend the cells with 500 μ L of the washing buffer and analyze by a flow cytometer. (Positive control for flow cytometry; transfectant)
Reference Data