United Kingdom (UK)
Recombinant monoclonal antibody to Progesterone Receptor. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 14B3.
UniProt Accession Number of Target Protein: P06401
Alternative Name(s) of Target: PR; Nuclear receptor subfamily 3 group C member 3
Immunogen: The original antibody was generated by immunizing rabbits with PR protein.
Specificity: The antibody is specific for PR protein. PR is a protein molecule, a cellular core receptor belonging to a steroidal superfamily, which can be bonded to the corresponding hormone to form a hormone-receptor complex, enabling hormones to exert their biological effects.
Application Notes: The specificity of the original format of the antibody was confirmed by ELISA analysis (affinity constant of 1.82 × 10^10 M-1). The antibody specifically detected PR protein in Western blot analysis. The antibody 14B3 and the commercial antibody 1E2 were used to stain breast cancer and endometrial. Results showed the antibody specificity is comparable to the commercial antibody. Further, the sensitivity of the antibody 14B3 was higher than the commercial antibody (CN113583132A).
Antibody first published in: PMID:
Western blot using anti-Progesterone Receptor antibody 14B3 (Ab03318). MCF7 cells lysate (35µg protein in RIPA buffer) was resolved on an SDS-PAGE gel and blots were probed with the chimeric rabbit version of 14B3 (Ab03318-23.0) at 0.01µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1 hour was used, and protein was detected by chemiluminescence.
Immunofluorescence staining of MCF7 cells with anti-Progesterone Receptor antibody 14B3 (Ab03318). Immunofluorescence analysis of paraformaldehyde fixed MCF7 cells, permeabilized with 0.15% Triton, on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of 14B3 (Ab03318-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing nuclear and cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, Ab03318-23.0, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.
Flow cytometry using anti-Progesterone Receptor antibody 14B3 (Ab03318). Paraformaldehyde fixed MCF7 cells permeabilized with 0.5% Triton were stained with the anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of 14B3 (Ab03318-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and the cells were analyzed using a FACSCanto flow-cytometer.
Western blot using anti-Progesterone Receptor antibody 14B3 (Ab03318). MCF7 cells lysate (35µg protein in RIPA buffer) was resolved on an SDS-PAGE gel and blots were probed with the chimeric rabbit version of 14B3 (Ab03318-23.0) at 0.01µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1 hour was used, and protein was detected by chemiluminescence.