Anti-RAP tag [PMab-2]

Description:

Recombinant monoclonal antibody to RAP tag. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma PMab-2.

Clone Name:

PMab-2

Published Application(s):

epitope tag, WB, ELISA, FC, IHC

Published Species Reactivity:

Rat

Products based on this clone:

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Ab00885-23.0 Anti-RAP tag [PMab-2]
Rat
Rabbit IgG
Purified
In Stock

Ab00885-1.1 Anti-RAP tag [PMab-2]
Rat
Mouse IgG1
Purified
In Stock

UniProt Accession Number of Target Protein: Q64294

Alternative Name(s) of Target: Podoplanin; E11 antigen epitope; Glycoprotein 38; Gp38; OTS-8; Ots8; RTI140; T1-alpha; T1A; Type I cell 40 kDa protein
Immunogen: This antibody was raised by immunising mice with 14-residue synthetic peptide rpp3851, which corresponds to amino acids 38-51 (GDDMVNPGLEDRIEC) of the platelet-aggregation-stimulating (PLAG) domain of rat podoplanin (PDPN). Spleen cells were then harvested, and fused to P3U1 cells to generate stable hybridomas.

Specificity: This antibody is specific for amino acids 38-51 (GDDMVNPGLEDRIEC) of the platelet-aggregation-stimulating (PLAG) domain of rat podoplanin (PDPN). PDPN is a is a type I transmembrane protein that interacts with the platelet receptor C-type lectin-like receptor-2 (CLEC-2). PDPN is expressed by a wide variety of cells, including podocytes, lymphatic endothelial cells and type I alveolar cells. PDPN is also upregulated on certain cancer cells, where it mediates tumour-induced platelet aggregation by binding to CLEC-2 on platelets

Application Notes: The specificity of this antibody for endogenous rat podoplanin (PDPN) has been confirmed in ELISA, Western blot and flow cytometry analysis (Oki et al, 2015), where PMab-2 was shown not to react with human and mouse podoplanin. Flow cytometry has also been used to demonstrate the high binding affinity of this antibody (KD of PMab-2 = 5.9 × 10−10 M) (Oki et al, 2015). In immunohistochemical analysis, this antibody is highly sensitive to rat PDPN, detecting type I lung alveolar cells, colon lymphatic endothelial cells, podocytes and Bowman's capsule epithelial cells at low concentration (Oki et al, 2015). This antibody possess high affinity and specificity for the RAP epitope tag (DMVNPGLEDRIE), and can, therefore, be used to detect proteins in Western blots, flow cytometry and sandwich enzyme-linked immunosorbent assays (ELISA), as well as for protein purification. (Fujii et al, 2017).

Antibody first published in: Oki et al Development of Sensitive Monoclonal Antibody PMab-2 Against Rat Podoplanin Monoclonal Antibodies in Immunodiagnosis and Immunotherapy. December 2015, 34(6): 396-403. PMID:26683179
Note on publication: Describes the original generation and characterisation of this antibody.

ELISA using Ab00885-1.1. Click on image for more details.

ELISA using PMab-2 (Ab00885) and Multi Tag protein 2. Binding curves of the mouse IgG1 chimeric version of the anti-RAP Tag antibody PMab-2 (Ab00885-1.1; yellow line) and isotype control (Ab00102-1.1, anti-fluorescein; orange line) to an ELISA plate coated with Multi Tag protein 2 (Pr00565, Absolute Antibody) at a concentration of 5 ug/ml. Ab02007 antibody was titrated on a 3-fold serial dilution starting at 10,000 ng/mL. Anti-mouse IgG was used as secondary antibody for detection at a dilution of 1:4,000.

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