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hca230z (Bio-Rad)

Name: hca230z
Brand: Bio-Rad
Rating: 92 (5 reviews)

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Bio-Rad hca230z

Hca230z, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/hca230z/product/Bio-Rad
Average 92 stars, based on 5 article reviews

hca230z - by Bioz Stars, 2026-02

92/100 stars

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1) Product Images from "Both enantiomers of β-aminoisobutyric acid BAIBA regulate Fgf23 via MRGPRD receptor by activating distinct signaling pathways in osteocytes"

Article Title: Both enantiomers of β-aminoisobutyric acid BAIBA regulate Fgf23 via MRGPRD receptor by activating distinct signaling pathways in osteocytes

Journal: Cell reports

doi: 10.1016/j.celrep.2024.114397

Figure Legend Snippet:

Techniques Used: Recombinant, Modification, Protease Inhibitor, Saline, Enzyme-linked Immunosorbent Assay, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Membrane, cAMP Assay, TaqMan Assay, Software

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Journal: Cell reports

Article Title: Both enantiomers of β-aminoisobutyric acid BAIBA regulate Fgf23 via MRGPRD receptor by activating distinct signaling pathways in osteocytes

doi: 10.1016/j.celrep.2024.114397

Figure Lengend Snippet:

Article Snippet: Sclerostin antibody AbD09097-h/m IgG2a , Bio-Rad , HCA230Z.

Techniques: Recombinant, Modification, Protease Inhibitor, Saline, Enzyme-linked Immunosorbent Assay, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Membrane, cAMP Assay, TaqMan Assay, Software

Journal: Cell reports

Article Title: Both enantiomers of β-aminoisobutyric acid BAIBA regulate Fgf23 via MRGPRD receptor by activating distinct signaling pathways in osteocytes

doi: 10.1016/j.celrep.2024.114397

Figure Lengend Snippet:

Article Snippet: Sclerostin antibody AbD09097-h/m IgG2a , Bio-Rad , HCA230Z.

Fab antibodies against murine and human sclerostin. Analysis of Fab antibodies derived from phage display using murine and human sclerostin as antigens. The binding affinities of these Fabs were determined using surface plasmon resonance (SPR), K D values and kinetic parameters were obtained using a set-up with the Fab antibody proteins as analytes (six different concentrations) and human and murine sclerostin as immobilized ligand. N.B. indicates no binding in the SPR experiment, which under the measurement conditions corresponds to a lower binding affinity boundary of less than 5 µM; n.d., not determined. Changes in bioactivity were measured by a Wnt-responsive reporter gene assay using HEK293TSA cells transfected with Wnt1. Indicated are the changes in IC 50 value upon addition of Fab antibody compared with cells treated with recombinant sclerostin proteins alone (means of at least two experiments).

Journal: Open Biology

Article Title: The sclerostin-neutralizing antibody AbD09097 recognizes an epitope adjacent to sclerostin's binding site for the Wnt co-receptor LRP6

doi: 10.1098/rsob.160120

Figure Lengend Snippet: Fab antibodies against murine and human sclerostin. Analysis of Fab antibodies derived from phage display using murine and human sclerostin as antigens. The binding affinities of these Fabs were determined using surface plasmon resonance (SPR), K D values and kinetic parameters were obtained using a set-up with the Fab antibody proteins as analytes (six different concentrations) and human and murine sclerostin as immobilized ligand. N.B. indicates no binding in the SPR experiment, which under the measurement conditions corresponds to a lower binding affinity boundary of less than 5 µM; n.d., not determined. Changes in bioactivity were measured by a Wnt-responsive reporter gene assay using HEK293TSA cells transfected with Wnt1. Indicated are the changes in IC 50 value upon addition of Fab antibody compared with cells treated with recombinant sclerostin proteins alone (means of at least two experiments).

Article Snippet: Among those, the sclerostin-neutralizing Fab AbD09097 (available as a full-length IgG as HCA230Z from AbD Serotec) or its affinity-maturated descendants might present the most interesting antibodies.

Techniques: Derivative Assay, Binding Assay, SPR Assay, Reporter Gene Assay, Transfection, Recombinant, Reporter Assay

AbD09097 binds to sclerostin's flexible loop. ( a ) Competition ELISA measuring the binding of selected Fab antibodies to immobilized sclerostin (human form, for AbD09096 and AbD09099 murine sclerostin) in the presence of the peptide tSOST-Δβ2ox (marked with A), which resembles a truncated sclerostin protein only comprising finger 1 and 2, and PFD-038IAM (marked with B and displayed in red), which mimics the linear form of sclerostin loop 2. The data are derived from a single experiment. ( b ) Competition ELISA measuring the binding of the Fab antibody AbD09097 to immobilized murine sclerostin in the presence of an increasing concentration of the peptides PFD-038IAM and tSOST-Δβ2ox. The binding of the Fab was determined by a secondary anti-Fab alkaline-phosphatase conjugate. The red and black data points indicate two separate experiments combined in one graph. ( c – f ) SPR sensograms are shown for the interaction of AbD09097 and AbD09096 with murine (mScl, c ), human (hScl, d ) and the murine sclerostin variants Δloop ( e ) and AlaLoop ( f ). The sclerostin proteins were immobilized on the chip surface. At time point zero AbD09097 was injected as analyte using six different concentrations (100, 75, 50, 25, 12.5 and 6.25 nM), curves for 75 nM are shown. At time point 200 s injection of the Fab protein was stopped and the chip surface was perfused with buffer. Fitted curves are shown as dashed lines. ( g – h ) SPR competition experiment for the Fabs AbD09096 ( g ) and AbD09097 ( h ). The left bars (marked with w/o) show the signal (in resonance unit, RU) measured at 200 s perfusion of 300 nM Fab fragment over a biosensor coated with 600 RU wild-type murine sclerostin. The middle bars (marked with mScl) show the signal obtained when 300 nM of the respective Fab antibody was perfused in the presence of 600 nM wild-type murine sclerostin. As sclerostin shows unspecific binding to the chip surface, the signal from perfusing 600 nM sclerostin without any Fab protein was subtracted from the raw signal obtained for co-injection of Fab and sclerostin. The right bars (marked with mScl Δloop) show the signal after co-injecting 300 nM Fab in the presence of 600 nM of the variant Scl Δloop, which lacks loop 2. Data processing included subtraction of an injection with 600 nM Scl Δloop to remove the effect of unspecific binding of sclerostin to the chip surface as stated above. Shown are mean RU values with s.d. from at least eight experiments. Means of the binding signal for AbD09096 alone and co-injected with sclerostin or the variant mScl Δloop differ significantly ( p < 0.0001), just as do the means of the binding signal for AbD09097 alone and co-injected with murine sclerostin ( p < 0.0001). Means of AbD09097 alone and co-injected with the variant mScl Δloop do not differ significantly ( p = 0.1166). Statistical results were obtained with an unpaired two-tailed Student's t -test.

Journal: Open Biology

Article Title: The sclerostin-neutralizing antibody AbD09097 recognizes an epitope adjacent to sclerostin's binding site for the Wnt co-receptor LRP6

doi: 10.1098/rsob.160120

Figure Lengend Snippet: AbD09097 binds to sclerostin's flexible loop. ( a ) Competition ELISA measuring the binding of selected Fab antibodies to immobilized sclerostin (human form, for AbD09096 and AbD09099 murine sclerostin) in the presence of the peptide tSOST-Δβ2ox (marked with A), which resembles a truncated sclerostin protein only comprising finger 1 and 2, and PFD-038IAM (marked with B and displayed in red), which mimics the linear form of sclerostin loop 2. The data are derived from a single experiment. ( b ) Competition ELISA measuring the binding of the Fab antibody AbD09097 to immobilized murine sclerostin in the presence of an increasing concentration of the peptides PFD-038IAM and tSOST-Δβ2ox. The binding of the Fab was determined by a secondary anti-Fab alkaline-phosphatase conjugate. The red and black data points indicate two separate experiments combined in one graph. ( c – f ) SPR sensograms are shown for the interaction of AbD09097 and AbD09096 with murine (mScl, c ), human (hScl, d ) and the murine sclerostin variants Δloop ( e ) and AlaLoop ( f ). The sclerostin proteins were immobilized on the chip surface. At time point zero AbD09097 was injected as analyte using six different concentrations (100, 75, 50, 25, 12.5 and 6.25 nM), curves for 75 nM are shown. At time point 200 s injection of the Fab protein was stopped and the chip surface was perfused with buffer. Fitted curves are shown as dashed lines. ( g – h ) SPR competition experiment for the Fabs AbD09096 ( g ) and AbD09097 ( h ). The left bars (marked with w/o) show the signal (in resonance unit, RU) measured at 200 s perfusion of 300 nM Fab fragment over a biosensor coated with 600 RU wild-type murine sclerostin. The middle bars (marked with mScl) show the signal obtained when 300 nM of the respective Fab antibody was perfused in the presence of 600 nM wild-type murine sclerostin. As sclerostin shows unspecific binding to the chip surface, the signal from perfusing 600 nM sclerostin without any Fab protein was subtracted from the raw signal obtained for co-injection of Fab and sclerostin. The right bars (marked with mScl Δloop) show the signal after co-injecting 300 nM Fab in the presence of 600 nM of the variant Scl Δloop, which lacks loop 2. Data processing included subtraction of an injection with 600 nM Scl Δloop to remove the effect of unspecific binding of sclerostin to the chip surface as stated above. Shown are mean RU values with s.d. from at least eight experiments. Means of the binding signal for AbD09096 alone and co-injected with sclerostin or the variant mScl Δloop differ significantly ( p < 0.0001), just as do the means of the binding signal for AbD09097 alone and co-injected with murine sclerostin ( p < 0.0001). Means of AbD09097 alone and co-injected with the variant mScl Δloop do not differ significantly ( p = 0.1166). Statistical results were obtained with an unpaired two-tailed Student's t -test.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Derivative Assay, Concentration Assay, Injection, Variant Assay, Two Tailed Test

NMR mapping study to determine the binding epitope of the neutralizing Fab AbD09097. ( a ) Overlay of 1 H 15 N 2D HSQC spectra of the uniformly 15 N-labelled murine sclerostin variant ΔNC in the absence (black) and presence (red) of an equimolar amount of AbD09097. NMR signals of sclerostin affected by the binding of AbD09097 (i.e. visible either from changes in the chemical shift or from altered line width) are indicated. ( b ) Bar diagram highlighting the residues involved in binding of AbD09097 as observed by NMR mapping. P denotes proline residues which do not have an amide nitrogen–proton correlation, asterisk (*) denotes residues whose correlation signal could not be assigned unambiguously. ( c ) Van der Waal surface representation of sclerostin with the amino acid residues whose NMR signals were affected upon binding of AbD09097 highlighted in red. In the right panel, the sclerostin structure is rotated around the y -axis by 90°.

Journal: Open Biology

Article Title: The sclerostin-neutralizing antibody AbD09097 recognizes an epitope adjacent to sclerostin's binding site for the Wnt co-receptor LRP6

doi: 10.1098/rsob.160120

Figure Lengend Snippet: NMR mapping study to determine the binding epitope of the neutralizing Fab AbD09097. ( a ) Overlay of 1 H 15 N 2D HSQC spectra of the uniformly 15 N-labelled murine sclerostin variant ΔNC in the absence (black) and presence (red) of an equimolar amount of AbD09097. NMR signals of sclerostin affected by the binding of AbD09097 (i.e. visible either from changes in the chemical shift or from altered line width) are indicated. ( b ) Bar diagram highlighting the residues involved in binding of AbD09097 as observed by NMR mapping. P denotes proline residues which do not have an amide nitrogen–proton correlation, asterisk (*) denotes residues whose correlation signal could not be assigned unambiguously. ( c ) Van der Waal surface representation of sclerostin with the amino acid residues whose NMR signals were affected upon binding of AbD09097 highlighted in red. In the right panel, the sclerostin structure is rotated around the y -axis by 90°.

Techniques: Binding Assay, Variant Assay

Peptide replacement array analysis for sclerostin-neutralizing Fabs on all single mutant variants of peptide PARLLPNAIGRGKW representing residues Pro86 to Trp99 of loop 2 of murine sclerostin. The red line represents the mean signal height that is recorded for the base peptide. At each position along the sequence, a letter representing the substitution is drawn at the height of the signal intensity recorded for that particular substitution. One letter amino acid code was used for all amino acids except Cys, for which the number 2 was used representing a cysteine protected with acetamidomethyl (ACM) to prevent formation of disulfide bonds. For all three Fabs, AbD09097 ( a ), AbD12682 ( b ) and AbD12683 ( c ), the first eight residues can be replaced by any other tested amino acid without altering binding of the peptide. Exchange of the succeeding six amino acids, representing residues Ile94 to Trp99 of sclerostin, similarly but not identically affect binding of the peptide by the different Fab proteins.

Journal: Open Biology

Article Title: The sclerostin-neutralizing antibody AbD09097 recognizes an epitope adjacent to sclerostin's binding site for the Wnt co-receptor LRP6

doi: 10.1098/rsob.160120

Figure Lengend Snippet: Peptide replacement array analysis for sclerostin-neutralizing Fabs on all single mutant variants of peptide PARLLPNAIGRGKW representing residues Pro86 to Trp99 of loop 2 of murine sclerostin. The red line represents the mean signal height that is recorded for the base peptide. At each position along the sequence, a letter representing the substitution is drawn at the height of the signal intensity recorded for that particular substitution. One letter amino acid code was used for all amino acids except Cys, for which the number 2 was used representing a cysteine protected with acetamidomethyl (ACM) to prevent formation of disulfide bonds. For all three Fabs, AbD09097 ( a ), AbD12682 ( b ) and AbD12683 ( c ), the first eight residues can be replaced by any other tested amino acid without altering binding of the peptide. Exchange of the succeeding six amino acids, representing residues Ile94 to Trp99 of sclerostin, similarly but not identically affect binding of the peptide by the different Fab proteins.

Techniques: Mutagenesis, Sequencing, Binding Assay

Crystal structure analysis of the sclerostin-neutralizing Fab AbD09097. ( a ) Stereo view of a ribbon plot of the Fab AbD09097. The constant (C) and variable (V) domains of the light (subscript L) and heavy (subscript H) chain are indicated. The six complementarity-determining regions (CDRs) are marked with L and H for the light and heavy chain. ( b ) Van der Waals surface representation (stereo view) of the antigen-binding site (as in ( a ) but viewed from the top). The CDR loops are colour-coded as in ( a ). A deep pocket (marked by a white line) is formed in the centre of the antigen-binding site, which is limited by the CDRs 1 and 3 of the light and heavy chain (L1, H1, L3 and H3). A small part of the CDR2 of the light chain (L2) and the heavy chain (H2) also contributes to the pocket.

Journal: Open Biology

Article Title: The sclerostin-neutralizing antibody AbD09097 recognizes an epitope adjacent to sclerostin's binding site for the Wnt co-receptor LRP6

doi: 10.1098/rsob.160120

Figure Lengend Snippet: Crystal structure analysis of the sclerostin-neutralizing Fab AbD09097. ( a ) Stereo view of a ribbon plot of the Fab AbD09097. The constant (C) and variable (V) domains of the light (subscript L) and heavy (subscript H) chain are indicated. The six complementarity-determining regions (CDRs) are marked with L and H for the light and heavy chain. ( b ) Van der Waals surface representation (stereo view) of the antigen-binding site (as in ( a ) but viewed from the top). The CDR loops are colour-coded as in ( a ). A deep pocket (marked by a white line) is formed in the centre of the antigen-binding site, which is limited by the CDRs 1 and 3 of the light and heavy chain (L1, H1, L3 and H3). A small part of the CDR2 of the light chain (L2) and the heavy chain (H2) also contributes to the pocket.

Techniques: Binding Assay

The C-termini of the Fab light and heavy chain potentially mimic the interaction of sclerostin with the Fab AbD09097. ( a ) Crystal-lattice contacts (the second Fab molecule is indicated in light grey, being stacked on top of the other Fab molecule) between symmetry-related Fab molecules result in the interaction of the C-terminus of the light chain (aa N211–A215 and K191) with the CDRs L1, L3, H2 and H3 of the antibody. Similarly, the C-terminus of the heavy chain comprising mainly the residues of the thrombin cleavage site for the removal of the Myc and hexahistidine-tag (aa S217–G224) interacts with the deep pocket formed by the CDRs L1, L2, L3, H1 and H3. ( b ) Magnification (stereo view) of the interaction of the C-terminus of the light chain (carbon atoms coloured in magenta) with the binding site of the Fab. Selected hydrogen bonds are marked as magenta stippled lines. Residues of the light chain are coloured in dark green and labels contain a subscript L. Residues of the heavy chain are coloured in green and labels contain a subscript H. Numbering of the residues located in the CDRs follows the rules of Kabat. ( c ) As in ( b ) but rotated around the y -axis by 180° to show the interaction of the C-terminus of the heavy chain (carbon atoms coloured in cyan) with the antigen-binding site of the Fab. Two amino acid residues of the thrombin cleavage site, F219 and R223, deeply penetrate into the pocket. Arginine 223 forms multiple hydrogen bonds emanating from its main and side chain atoms to residues Y36 and D50 of the light chain and F96 and D101 of the heavy chain.

Journal: Open Biology

Article Title: The sclerostin-neutralizing antibody AbD09097 recognizes an epitope adjacent to sclerostin's binding site for the Wnt co-receptor LRP6

doi: 10.1098/rsob.160120

Figure Lengend Snippet: The C-termini of the Fab light and heavy chain potentially mimic the interaction of sclerostin with the Fab AbD09097. ( a ) Crystal-lattice contacts (the second Fab molecule is indicated in light grey, being stacked on top of the other Fab molecule) between symmetry-related Fab molecules result in the interaction of the C-terminus of the light chain (aa N211–A215 and K191) with the CDRs L1, L3, H2 and H3 of the antibody. Similarly, the C-terminus of the heavy chain comprising mainly the residues of the thrombin cleavage site for the removal of the Myc and hexahistidine-tag (aa S217–G224) interacts with the deep pocket formed by the CDRs L1, L2, L3, H1 and H3. ( b ) Magnification (stereo view) of the interaction of the C-terminus of the light chain (carbon atoms coloured in magenta) with the binding site of the Fab. Selected hydrogen bonds are marked as magenta stippled lines. Residues of the light chain are coloured in dark green and labels contain a subscript L. Residues of the heavy chain are coloured in green and labels contain a subscript H. Numbering of the residues located in the CDRs follows the rules of Kabat. ( c ) As in ( b ) but rotated around the y -axis by 180° to show the interaction of the C-terminus of the heavy chain (carbon atoms coloured in cyan) with the antigen-binding site of the Fab. Two amino acid residues of the thrombin cleavage site, F219 and R223, deeply penetrate into the pocket. Arginine 223 forms multiple hydrogen bonds emanating from its main and side chain atoms to residues Y36 and D50 of the light chain and F96 and D101 of the heavy chain.

Techniques: Binding Assay

Affinity maturation of the Fab AbD09097. Analysis of Fab antibodies derived from affinity maturation of the ancestor Fab AbD09097. The Fab AbD12533 is a bivalent AbD09097, in which two AbD09097 were fused by a helix-turn-helix motif [ <xref ref-type=

65 ]. Fab specificity was screened using an ELISA employing a panel of control proteins, and murine and human sclerostin. The numbers represent signal/noise ratios (fold background). The binding affinity of these Fabs were determined using surface plasmon resonance (SPR), K D values and kinetic parameters were obtained using a set-up with the Fab antibody proteins as analytes (six different concentrations) and human and murine sclerostin as immobilized ligand. Fold change represents the affinity enhancement compared with the binding affinity of the ancestor Fab AbD09097. Species-specificity was calculated from ." width="100%" height="100%">

Journal: Open Biology

Article Title: The sclerostin-neutralizing antibody AbD09097 recognizes an epitope adjacent to sclerostin's binding site for the Wnt co-receptor LRP6

doi: 10.1098/rsob.160120

Figure Lengend Snippet: Affinity maturation of the Fab AbD09097. Analysis of Fab antibodies derived from affinity maturation of the ancestor Fab AbD09097. The Fab AbD12533 is a bivalent AbD09097, in which two AbD09097 were fused by a helix-turn-helix motif [ 65 ]. Fab specificity was screened using an ELISA employing a panel of control proteins, and murine and human sclerostin. The numbers represent signal/noise ratios (fold background). The binding affinity of these Fabs were determined using surface plasmon resonance (SPR), K D values and kinetic parameters were obtained using a set-up with the Fab antibody proteins as analytes (six different concentrations) and human and murine sclerostin as immobilized ligand. Fold change represents the affinity enhancement compared with the binding affinity of the ancestor Fab AbD09097. Species-specificity was calculated from .

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Control, Binding Assay, SPR Assay

Model of the sclerostin–AbD09097 complex. On the basis of the C-termini of the heavy and light chains of a symmetry-related Fab molecule in the crystal lattice of the AbD09097 crystal a theoretical model was built to show the loop 2 of human sclerostin comprising Asn92 to Ser103 bound into the antigen-binding crevice of AbD09097. The carbon atoms of the heavy and light chains of AbD09097 are coloured in light and dark grey, respectively. The carbon atoms of the sclerostin loop are shown in green. Potential hydrogen bonds between the sclerostin loop and Fab are indicated by stippled lines in magenta. ( a ) Top down view of the Fab-binding cleft. ( b ) As in ( a ) but rotated 90° clockwise around the x -axis.

Journal: Open Biology

Article Title: The sclerostin-neutralizing antibody AbD09097 recognizes an epitope adjacent to sclerostin's binding site for the Wnt co-receptor LRP6

doi: 10.1098/rsob.160120

Figure Lengend Snippet: Model of the sclerostin–AbD09097 complex. On the basis of the C-termini of the heavy and light chains of a symmetry-related Fab molecule in the crystal lattice of the AbD09097 crystal a theoretical model was built to show the loop 2 of human sclerostin comprising Asn92 to Ser103 bound into the antigen-binding crevice of AbD09097. The carbon atoms of the heavy and light chains of AbD09097 are coloured in light and dark grey, respectively. The carbon atoms of the sclerostin loop are shown in green. Potential hydrogen bonds between the sclerostin loop and Fab are indicated by stippled lines in magenta. ( a ) Top down view of the Fab-binding cleft. ( b ) As in ( a ) but rotated 90° clockwise around the x -axis.

Techniques: Binding Assay

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1) Product Images from "Both enantiomers of β-aminoisobutyric acid BAIBA regulate Fgf23 via MRGPRD receptor by activating distinct signaling pathways in osteocytes"

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