Journal: Frontiers in Immunology

Article Title: C1q and Mannose-Binding Lectin Interact with CR1 in the Same Region on CCP24-25 Modules

doi: 10.3389/fimmu.2018.00453

Figure Lengend Snippet: Kinetic and equilibrium dissociation constants for the binding of C1q to immobilized soluble CR1 (sCR1) and CR1 CCP22-30.

Article Snippet: Recombinant soluble human CR1 (sCR1) was purchased from R&D Systems Europe.

Techniques: Binding Assay

Journal: Frontiers in Immunology

Article Title: C1q and Mannose-Binding Lectin Interact with CR1 in the Same Region on CCP24-25 Modules

doi: 10.3389/fimmu.2018.00453

Figure Lengend Snippet: Localization of the C1q binding sites in CR1 CCP22-30 using deletion fragments. (A) Schematic view of the different fragments of CR1 CCP22-30. Potential N-glycosylations are represented by open circles ○. (B) Interaction of C1q with immobilized CR1 CCP22-30 fragments analyzed by ELISA. C1q (10 µg/ml in PBS) was added to microtiter plate coated with 3.4 pmol of each CCP22-30 fragment as described in Section “ .” After 1.5 h incubation at room temperature, bound C1q was detected with rabbit antibodies recognizing C1q and HRP secondary antibodies. Data are presented as the mean ± SE of four individual experiments. (**), Student’s t -test values p < 0.01 of C1q binding to each fragment compared to CCP22-30.

Article Snippet: Recombinant soluble human CR1 (sCR1) was purchased from R&D Systems Europe.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation

Journal: Frontiers in Immunology

Article Title: C1q and Mannose-Binding Lectin Interact with CR1 in the Same Region on CCP24-25 Modules

doi: 10.3389/fimmu.2018.00453

Figure Lengend Snippet: Complement receptor type 1 (CR1) variants produced to study the CCP24-25 module pair interaction properties. (A) Schematic view of CCP24-25 variants produced in eukaryotic cells. Potential N-glycosylations are represented by open circles ○. (B) SDS-PAGE analysis of 4 µg of the CR1variants under reducing conditions. The positions of the molecular weight markers (expressed in kilodaltons) are indicated. A full scan of the original gel is provided in Figure S1 in Supplementary Material .

Article Snippet: Recombinant soluble human CR1 (sCR1) was purchased from R&D Systems Europe.

Techniques: Produced, SDS Page, Molecular Weight

Journal: Frontiers in Immunology

Article Title: C1q and Mannose-Binding Lectin Interact with CR1 in the Same Region on CCP24-25 Modules

doi: 10.3389/fimmu.2018.00453

Figure Lengend Snippet: Kinetic analysis of the interaction of C1q and mannose-binding lectin (MBL) with CR1 CCP24-25. C1q (A) and MBL (B) , at indicated concentrations, were injected in 50 mM triethanolamine-HCl (TEA), 145 mM NaCl, 0.05% surfactant P20, pH 7.4, on immobilized CR1 CCP24-25 (1,000 RU). The buffer was supplemented with 3 mM EDTA for MBL binding. Fits are shown as dotted lines and were obtained by global fitting of the data using a 1:1 Langmuir-binding model. The kinetic constants obtained are framed on the top of each sensorgramm.

Article Snippet: Recombinant soluble human CR1 (sCR1) was purchased from R&D Systems Europe.

Techniques: Binding Assay, Injection

Journal: Frontiers in Immunology

Article Title: C1q and Mannose-Binding Lectin Interact with CR1 in the Same Region on CCP24-25 Modules

doi: 10.3389/fimmu.2018.00453

Figure Lengend Snippet: Interaction of C1q and mannose-binding lectin (MBL) with immobilized CR1 CCP22-30 and CR1 ΔCCP24-25. The binding curves were obtained in single cycle mode by injecting increasing concentrations of C1q (A) or MBL (B) on immobilized CR1 CCP22-30 (3,000 RU) or CR1 ΔCCP24-25 (2,200 RU). C1q (0.25, 0.5, 1, 2, and 4 nM) was injected at 20 µl/min for 180 s in 50 mM triethanolamine-HCl (TEA), 150 mM NaCl, 1 mM CaCl 2 , 0.05% surfractant P20, pH 7.4. MBL (1, 2, 4, 8, and 16 nM) was injected in the same conditions except that the buffer contained 3 mM EDTA instead of CaCl 2 .

Article Snippet: Recombinant soluble human CR1 (sCR1) was purchased from R&D Systems Europe.

Techniques: Binding Assay, Injection

Journal: Frontiers in Immunology

Article Title: C1q and Mannose-Binding Lectin Interact with CR1 in the Same Region on CCP24-25 Modules

doi: 10.3389/fimmu.2018.00453

Figure Lengend Snippet: Elongated shape of CCP24-25, associated to a possible interpretative model. (A) Small-angle X-ray scattering (SAXS) pair distance distribution, (B) SAXS dimensionless Kratky plot, which shows elongation and remaining flexibility. (C) Fit of the model [shown in panels (D,E) ] to the experimental data. (D) Top and (E) side views of an ab initio envelope computed with GASBOR. A Coral-derived model of CCP24-25 is shown inside to illustrate how the shape corresponds to two CCP modules. This interpretative model is used to illustrate the positions on each module of glycosylation sites (green), rs 4844609 SNP (magenta), Knops groups variants (cyan), acidic clusters (red) initially suggested as potential MBL binding sites, and the other acidic residues (gray and black). D1529 is the homologous counterpart of D1076 in CR1 CCP17 (black). The two flexible carbohydrates included in the model are only illustrated in the side view.

Article Snippet: Recombinant soluble human CR1 (sCR1) was purchased from R&D Systems Europe.

Techniques: Derivative Assay, Glycoproteomics, Binding Assay

Journal: Frontiers in Immunology

Article Title: C1q and Mannose-Binding Lectin Interact with CR1 in the Same Region on CCP24-25 Modules

doi: 10.3389/fimmu.2018.00453

Figure Lengend Snippet: Localization of the binding site of soluble CR1 on C1q. (A) Comparative binding of C1q and its CLF and GR on complement receptor type 1 (CR1) CCP22-30 analyzed by surface plasmon resonance (SPR). The CR1 CCP22-30 fragment was immobilized on CM5 sensor chips (4,500 RU) and 2 nM of C1q or CLF and 12 nM of GR were injected over the surfaces as described in Section “ .” (B) MBL-associated serine protease (MASP)-3 competition of the binding of C1q to CR1 CCP22-30 analyzed by SPR. C1q (2 nM) was incubated at room temperature for 20 min in the absence or presence of recombinant MASP-3 at indicated molar ratios and injected over immobilized CR1 CCP22-30 (4,500 RU). No binding was observed when MASP-3 alone (20 nM) was injected.

Article Snippet: Recombinant soluble human CR1 (sCR1) was purchased from R&D Systems Europe.

Techniques: Binding Assay, SPR Assay, Injection, Incubation, Recombinant

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Sulforaphane suppresses the growth of triple-negative breast cancer stem-like cells in vitro and in vivo

doi: 10.1158/1940-6207.CAPR-18-0241

Figure Lengend Snippet: Selected validation of Nanostring gene expression analysis by qRT-PCR. Four genes including CR1, FOXD3, WNT3, and NOTCH4 are shown here as representatives for amplification. Data are the average of 8 samples from SFN pre-treated group and 8 samples from saline control. All experiments were conducted in duplicates. *: P < 0.05 as compared to the control animals.

Article Snippet: CR1 (R&D Systems, Cat# 145-CR/CF), Nodal (R&D Systems, Cat# 3218-ND-024/CF), GRP78 (Abcam, Cat# ab78432), and Alk4 (Creative BioMart, Cat# ACVR1B-645H,) were purchased from the indicated vendors.

Techniques: Biomarker Discovery, Gene Expression, Quantitative RT-PCR, Amplification, Saline, Control

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Sulforaphane suppresses the growth of triple-negative breast cancer stem-like cells in vitro and in vivo

doi: 10.1158/1940-6207.CAPR-18-0241

Figure Lengend Snippet: ELISA assessment of CR1 binding to solid phase BPs including Nodal, GRP78, and Alk4 in the absence (-S) or presence (+S) of 100µM of SFN. CR1 binding to individual BPs in the absence of SFN (-S) condition was normalized to 100%. Percentage reaction induced CR1 binding inhibition was then determined using the 100% reference point for respective BPs. Data represents average values determined from three separate studies with samples run in octuplicate.

Article Snippet: CR1 (R&D Systems, Cat# 145-CR/CF), Nodal (R&D Systems, Cat# 3218-ND-024/CF), GRP78 (Abcam, Cat# ab78432), and Alk4 (Creative BioMart, Cat# ACVR1B-645H,) were purchased from the indicated vendors.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Inhibition

Journal: Cancers

Article Title: Human Cripto-1 and Cripto-3 Protein Expression in Normal and Malignant Settings That Conflicts with Established Conventions

doi: 10.3390/cancers16213577

Figure Lengend Snippet: Amino acid (AA) sequence comparison of human CR1 (Accession No. AAH67844.1) and human CR3 (Accession No. AAG49539.1). The “*” indicates AA differences between CR1 and CR3. Yellow highlight = signal peptide. Fuchsia highlight = Abcam anti-CR1 MoAb ab108391 binding site (Leu44-Arg66). Red highlight = EFG-like domain (Nodal binding). Blue highlight = Abcam anti-CR1 MoAb ab133236 binding site (Cys97-Glu113). Purple highlight = CFC domain (GRP78/Alk4 binding). Green highlight = Glycoslylphosphatidylinositol (GPI) linkage domain. Double-headed arrow indicates the site of GPI-phospholipase D cleavage, resulting in the formation of soluble protein products. CR3A = peptide immunogen (srgdlafrdds) conjugated to KLH and used to generate anti-CR3 mouse MoAbs. CR3B = negative control peptide (srgylafrdds). CR1A = peptide immunogen (qrvppmgiqhs) conjugated to KLH and used to generate anti-CR1 mouse MoAbs. CR1B = negative control peptide (qrvlpmgiqhs).

Article Snippet: The resulting standard curve for GenScript recombinant human CR1 captured on solid-phased rabbit anti-CR1 was detected with anti-CR1 NCI 5G1-1 ( A).

Techniques: Sequencing, Comparison, Binding Assay, Negative Control

Journal: Cancers

Article Title: Human Cripto-1 and Cripto-3 Protein Expression in Normal and Malignant Settings That Conflicts with Established Conventions

doi: 10.3390/cancers16213577

Figure Lengend Snippet: GenScript Biotech Corporation mouse anti-CR1 and anti-CR3 generation data.

Article Snippet: The resulting standard curve for GenScript recombinant human CR1 captured on solid-phased rabbit anti-CR1 was detected with anti-CR1 NCI 5G1-1 ( A).

Techniques: Clone Assay

Journal: Cancers

Article Title: Human Cripto-1 and Cripto-3 Protein Expression in Normal and Malignant Settings That Conflicts with Established Conventions

doi: 10.3390/cancers16213577

Figure Lengend Snippet: GenScript Biotech Corporation mouse MoAb production data and resulting titration curves for NCI final candidate choice. ( A ) Titration curve for anti-CR1 MoAb 5G1-1 evaluated on the following, solid-phased: Blue—R&D Systems recombinant human CR1, fuchsia—CR1A peptide immunogen, green—CR1B negative control peptide, gray—R&D Systems recombinant mouse CR1, yellow—R&D Systems recombinant human cryptic, orange—MyBioSource recombinant human CR3, black—CR3A peptide immunogen, and red—CR3B negative control peptide. ( B ) Titration curve for anti-CR3 5G11-2 evaluated on the following, solid-phased: Orange—MyBioSource recombinant human CR3, black—CR3A peptide immunogen, red—CR3B negative control peptide, blue—R&D Systems recombinant human CR1, fuchsia—CR1A peptide immunogen, green—CR1B negative control peptide, gray—R&D Systems recombinant mouse CR1, and yellow—R&D Systems recombinant human cryptic.

Article Snippet: The resulting standard curve for GenScript recombinant human CR1 captured on solid-phased rabbit anti-CR1 was detected with anti-CR1 NCI 5G1-1 ( A).

Techniques: Titration, Recombinant, Negative Control

Journal: Cancers

Article Title: Human Cripto-1 and Cripto-3 Protein Expression in Normal and Malignant Settings That Conflicts with Established Conventions

doi: 10.3390/cancers16213577

Figure Lengend Snippet: Biacore/surface plasmon resonance analysis of anti-CR1 NCI 5G1-1 MoAb and anti-CR3 NCI 5G11-2 MoAb binding to recombinant human CR1 or CR3. ( A ) NCI 5G1-1 MoAb (blue) diluted at 0.8, 4, 20, 100, and 500 nM versus NCI 5G11-2 (orange) at the same test concentrations measured interaction with immobilized CR1. ( B ) NCI 5G11-2 MoAb (orange) diluted at 0.8, 4, 20, 100, and 500 nM versus NCI 5G1-1 (blue) at the same test concentrations measured interaction with immobilized CR3.

Article Snippet: The resulting standard curve for GenScript recombinant human CR1 captured on solid-phased rabbit anti-CR1 was detected with anti-CR1 NCI 5G1-1 ( A).

Techniques: SPR Assay, Binding Assay, Recombinant

Journal: Cancers

Article Title: Human Cripto-1 and Cripto-3 Protein Expression in Normal and Malignant Settings That Conflicts with Established Conventions

doi: 10.3390/cancers16213577

Figure Lengend Snippet: Dilution of test antibodies giving 50% deflection of binding titration curve via solid-phase ELISA.

Article Snippet: The resulting standard curve for GenScript recombinant human CR1 captured on solid-phased rabbit anti-CR1 was detected with anti-CR1 NCI 5G1-1 ( A).

Techniques: Binding Assay, Titration, Enzyme-linked Immunosorbent Assay, Control

Journal: Cancers

Article Title: Human Cripto-1 and Cripto-3 Protein Expression in Normal and Malignant Settings That Conflicts with Established Conventions

doi: 10.3390/cancers16213577

Figure Lengend Snippet: IHC staining of human normal/tumor tissue. ( A ) Lung IHC analysis: ( A1 ) Normal lung, airway epithelium −CR3/+CR3, ( A2 ) Squamous CA, +CR1/+CR3, ( A3 ) Adenosquamous CA, tumor vascular endothelium +CR1/−CR3 (Anatomical Separation), tumor cells −CR1/+CR3, ( A3′ ) Enlargement of +CR1 blood vessels—black arrowheads, ( A4 ) Adenocarcinoma, +CR1/+CR3, ( A5 ) Adenocarcinoma, tumor vascular endothelium +CR1/−CR3 (Anatomical Separation), and tumor cells −CR1/+CR3, and ( A5′ ) Enlargement of +CR1 blood vessels—black arrowheads. ( B ) Colon IHC analysis: ( B1 ) Normal colon, cryptic epithelium +CR1/+CR3, ( B2 ) Adenocarcinoma, tumor vascular endothelium +CR1/−CR3 (Anatomical Separation Variant), tumor cells +CR1/+CR3, ( B2′ ) enlargement of +CR1 blood vessels—black arrowheads, ( B3 ) Adenocarcinoma, tumor cells −CR1/+CR3, ( B4 ) Adenocarcinoma, tumor vascular endothelium +CR1/−CR3 (Anatomical Separation Variant), tumor cells +CR1/+CR3, ( B4′ ) enlargement of +CR1 blood vessels—black arrowheads, and ( B5 ) Adenocarcinoma, tumor cells −CR1/+CR3. ( C ) Breast IHC analysis: ( C1 ) Normal breast, lobular ductal epithelium +CR1/+CR3, ( C2 – C5 ) Invasive ductal adenocarcinoma, tumor cells +CR1/+CR3. ( D ) Ovary IHC analysis: ( D1 ) Normal ovary mesothelium +CR1/+CR3, ( D2 ) Normal pellucida of the primary follicles +CR1/+CR3, ( D3 ) Normal cells of the inner theca +CR1/+CR3, ( D4 ) Endometrioid adenocarcinoma −CR1/+CR3, ( D5 ) Metastatic adenocarcinoma +CR1/+CR3, ( D6 ) Dysgerminoma −CR1/+CR3, and ( D7 ) High-grade serous carcinoma +CR1/+CR3. ( E ) Prostate IHC analysis: ( E1 ) Normal prostate −CR1/−CR3, ( E2 ) Normal-looking tissue adjacent to adenocarcinoma +CR1/+CR3, possibly representing a premalignant lesion, ( E3 ) Adenocarcinoma +/−CR1/+CR3, ( E4 ) Adenocarcinoma +CR1/+CR3, and ( E5 ) Adenocarcinoma −CR1/+CR3. Photographs taken at 20× magnification and size bar constant for all figures.

Article Snippet: The resulting standard curve for GenScript recombinant human CR1 captured on solid-phased rabbit anti-CR1 was detected with anti-CR1 NCI 5G1-1 ( A).

Techniques: Immunohistochemistry, Variant Assay

Journal: Cancers

Article Title: Human Cripto-1 and Cripto-3 Protein Expression in Normal and Malignant Settings That Conflicts with Established Conventions

doi: 10.3390/cancers16213577

Figure Lengend Snippet: IHC detection of CR1/CR3 in human tissue and ranking of TMN_N, TMN_M, grade, stage, and progesterone receptor expression based on staining intensity. ( A ) Discriminate TMN_N0 from TMN_N1 in prostate cancer as scored by CR1 staining intensity. ( B ) Ranking of TMN_M0 vs. 5TMN_M1 in prostate cancer as revealed by CR1 staining. ( C ) Stage values for prostate cancer tracks with CR1 staining. ( D ) Discriminate colon mucinous adenocarcinoma grades +, ++, & +++ by CR3 staining. ( E ) Ranking of colon adenocarcinoma grades +, ++, and +++ based on CR3 staining. ( F ) Progesterone receptor score tracks with CR3 staining in breast cancer. Photographs taken at 20× magnification, and size bar constant for all figures. Dots indicate outliers.

Article Snippet: The resulting standard curve for GenScript recombinant human CR1 captured on solid-phased rabbit anti-CR1 was detected with anti-CR1 NCI 5G1-1 ( A).

Techniques: Expressing, Staining

Journal: Cancers

Article Title: Human Cripto-1 and Cripto-3 Protein Expression in Normal and Malignant Settings That Conflicts with Established Conventions

doi: 10.3390/cancers16213577

Figure Lengend Snippet: CR1 versus CR3 expression in human tumor cell lines as revealed by Western blot (WB) analysis. Note: CR1 and CR3 band intensity values normalized to respective GAPDH levels for the analyzed sample. Identification of human tumor cell lines and negative control. MCF7—breast CA, MDA-MB231—triple-negative breast CA, HepG2—hepatocellular CA, HT29—colorectal adeno CA, A549—bronchioloalveolar CA, hmVECs—immortalized endothelial cells. ( A ) Combined WB data for CR1 versus CR3 and corresponding housekeeping GAPDH loading standard. ( B ) Quantitative analysis of relative CR1 versus CR3 expression levels in cell lysates normalized to GAPDH loading control. Uncropped blots are shown in .

Article Snippet: The resulting standard curve for GenScript recombinant human CR1 captured on solid-phased rabbit anti-CR1 was detected with anti-CR1 NCI 5G1-1 ( A).

Techniques: Expressing, Western Blot, Negative Control, Control

Journal: Cancers

Article Title: Human Cripto-1 and Cripto-3 Protein Expression in Normal and Malignant Settings That Conflicts with Established Conventions

doi: 10.3390/cancers16213577

Figure Lengend Snippet: CR1/CR3 capture/quantitative ELISA for evaluation of soluble CR1/CR3 proteins in sera. ( A ) Representative CR1 capture/quantitative ELISA standard curve along with respective inserted linear equation graph used to determine CR1 values [ng/50 mL/well] of unknown serum sample. ( B ) Representative CR3 capture/quantitative ELISA standard curve along with respective inserted linear equation graph used to determine CR3 values [ng/50 mL/well] of unknown serum sample. ( C ) CR1 levels [ng/mL] identified in normal female donors and breast cancer patient sera. ( D ) CR3 levels [ng/mL] identified in normal female donors and breast cancer patient sera.

Article Snippet: The resulting standard curve for GenScript recombinant human CR1 captured on solid-phased rabbit anti-CR1 was detected with anti-CR1 NCI 5G1-1 ( A).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Cancers

Article Title: Human Cripto-1 and Cripto-3 Protein Expression in Normal and Malignant Settings That Conflicts with Established Conventions

doi: 10.3390/cancers16213577

Figure Lengend Snippet: CR1/CR3 interaction with solid-phased established CR1-binding proteins (Nodal, GRP78, and Alk4). To validate our assay was working correctly, we utilized positive control solid-phased antibodies that recognized antigenic epitopes on CR1/CR3 distal from NCI MoAb binding sites. These included the Abcam anti-CR1 N-terminal target (ab108391) = aNTCR1 and the anti-CR1 EGF-like domain target (ab133236) = aEGFCR1. The interaction of CR1 or CR3 to solid-phased Nodal, GRP78, or Alk4 proteins was detected with, respectively, anti-CR1 NCI 5G1-1 or anti-CR3 NCI 5G11-2 MoAbs. ( A ) CR1 interaction with solid-phased binding proteins or controls in the presence or absence of equimolar CR3. ( B ) CR3 interaction with solid-phased binding proteins or controls in the presence or absence of equimolar CR1. ( C ) CR1 percent binding inhibition with equimolar CR3. ( D ) CR3 percent binding inhibition with equimolar CR1. All target proteins were solid-phased at [50 ng/50 μL/well] overnight and blocked with HFBTS.

Article Snippet: The resulting standard curve for GenScript recombinant human CR1 captured on solid-phased rabbit anti-CR1 was detected with anti-CR1 NCI 5G1-1 ( A).

Techniques: Binding Assay, Positive Control, Inhibition

Journal: PLoS ONE

Article Title: Uptake and Presentation of Myelin Basic Protein by Normal Human B Cells

doi: 10.1371/journal.pone.0113388

Figure Lengend Snippet: PBMCs from healthy donors were incubated with or without 30 µg/ml MBP in medium containing normal autologous serum (30% v/v), or in pure medium. The resulting deposition of C3 and C1q on B cells was measured by flow cytometry after 5 min incubation (N = 3). Representative histogram plots show A) C3-deposition, and B) C1q-deposition on B cells. C) The binding of MBP was assessed using biotinylated MBP as probe and subsequent staining with streptavidin-PE. Blockade of CR1 or CR2 was achieved by pre-incubation of PBMCs with mAb3D9 and polyclonal sheep anti-human CR2 respectively. Monoclonal anti-glycophorin (GP)-A was used as negative control. D) Mean fluorescence intensity (MFI) values of 5–6 experiments are shown; background values (of samples with no MBP added) have been subtracted. Bars and error bars represent means and SEM. **p<0.01, ***p<0.001.

Article Snippet: Murine anti-human CR1 IgG1 antibody (mAb3D9) was kindly donated by Dr John O'Shea (Frederick Cancer Research and Development Center, Frederick, MD, USA), and polyclonal sheep anti-human CR2 was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Incubation, Flow Cytometry, Binding Assay, Staining, Negative Control, Fluorescence

Journal: PLoS ONE

Article Title: Uptake and Presentation of Myelin Basic Protein by Normal Human B Cells

doi: 10.1371/journal.pone.0113388

Figure Lengend Snippet: PBMCs from healthy HLA-DR15+ donors were incubated for 18 h with MBP in media containing normal serum. Cells were stained with FITC anti-CD19 and biotinylated MK16, followed by streptavidin-PE. (A) The binding of MK16 at different serum concentrations is shown as mean fluorescence (MFI) values normalised to that of 10% serum, (N = 4). B) Before addition of serum (30% v/v), different concentrations of the complement inhibitory compound sodium polyanethole sulphonate (SPS) were added. MFI values are shown, normalised to samples without SPS, (N = 6). (C) The PBMCs were pre-incubated with the anti-CR1 mAb3D9 or polyclonal sheep anti-human CR2, or both, before addition of serum (30% v/v) and MBP. Anti-glycophorin (GP)-A was used as negative control. Data are shown as means±SEM, (N = 4–6). **p<0.01.

Article Snippet: Murine anti-human CR1 IgG1 antibody (mAb3D9) was kindly donated by Dr John O'Shea (Frederick Cancer Research and Development Center, Frederick, MD, USA), and polyclonal sheep anti-human CR2 was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Incubation, Staining, Binding Assay, Fluorescence, Negative Control