United Kingdom (UK)
Recombinant monoclonal antibody to Asialoglycoprotein receptor. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the phage display antibody C2.
UniProt Accession Number of Target Protein: P07306
Alternative Name(s) of Target: ASGPR; ASGP-R; ASGR1; C-Type Lectin Domain Family 4 Member H1; Hepatic Lectin H1; CLEC4H1; HL-1;
Immunogen: A human non-immune phage antibody library was screened with the recombinant carbohydrate recognition domain of rCRDH1 (the H1 subunit of human asialoglycoprotein receptor). Anti-rCRDH1 phage clones were obtained after 4 rounds of screening with rCRDH1-coated immunotubes and single positive colonies were further selected with the expressed thioredoxin-His-S tag of the wild-type pET32c. Specific anti-rCRDH1 phage clones were transfected into E. coli HB2151 and induced for secreted expression of scFv antibody.
Specificity: C2 has been found to recognise denatured and native forms of asialoglycoprotein receptor.
Application Notes: C2 has been utilised in Western Blot and immunohistochemical experiments in order to establish its ability to bind to denatured ASGPR (Cao 2006). Since C2 is able to recognise denatured and native forms of the ASGPR, the antibody could potentially be used to target molecules for gene therapy of hepatocellular carcinoma or other liver diseases (Cao 2006).
Antibody first published in:
Cao et al. Characterization of a single-chain variable fragment (scFv) antibody directed against the human asialoglycoprotein receptor Biotechnol Appl Biochem. 2006 May;44(Pt 2):65-72. PMID:16396634
Note on publication:
Describes the generation of the C2 scFv and its potential application for targeting molecules for liver diseases, in addition to the advantages of using scFvs to give greater targeting specificity and better tumour penetration.
Immunofluorescence staining of HepG2 cells with anti-Asialoglycoprotein receptor (Ab00955) C2 Immunofluorescence analysis of paraformaldehyde fixed HepG2 cells stained with the chimeric rabbit IgG version of C2 (Ab00955-23.0) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00955-23.0, 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.
Western Blot using Anti-Asialoglycoprotein receptor antibody C2 (Ab00955). HepG2 cell lysate (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of C2 (Ab00955-23.0) at 1µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Flow cytometry using the Anti-Asialoglycoprotein receptor antibody C2 (Ab00955). Paraformaldehyde fixed HepG2 cells permeabilized with 0.5% Triton were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of C2 (Ab00955-23.0, 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 cells analyzed using a FACSCanto flow-cytometer.
Immunofluorescence staining of HepG2 cells with anti-Asialoglycoprotein receptor (Ab00955) C2 Immunofluorescence analysis of paraformaldehyde fixed HepG2 cells stained with the chimeric rabbit IgG version of C2 (Ab00955-23.0) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00955-23.0, 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.