cd21/cd35 antibody, anti-mouse, reafinity  (Miltenyi Biotec)

Journal: Analytical Chemistry

Article Title: Early Detection of Multiple Sclerosis Using Spectroscopic Epstein–Barr Virus Biosensors Based on Infection Mechanism Mimicry

doi: 10.1021/acs.analchem.5c03964

Figure Lengend Snippet: SEM (A–C) and AFM (D–F) images of (A, D) CD21@AuNP, (B,E) BSA@CD21@AuNP, and (C, F) gp350@BSA@CD21@AuNP.

Article Snippet: Recombinant human CD21 and recombinant EBV gp350 proteins were purchased from Sino Biological Inc. All reagents and chemicals employed in this study were analytical grade and used without further purification.

Techniques:

Journal: Analytical Chemistry

Article Title: Early Detection of Multiple Sclerosis Using Spectroscopic Epstein–Barr Virus Biosensors Based on Infection Mechanism Mimicry

doi: 10.1021/acs.analchem.5c03964

Figure Lengend Snippet: (A) FTIR and (B) UV spectra for the characterization of the developed colorimetric EBV biosensor. Step-by-step FTIR and UV spectra of the biosensor during gp350 detection: (a) 2.1 mL of AuNPs; (b) 2.1 mL of AuNPs + 400 μL of CD21 at 20 ng·mL –1 ; (c) 2.1 mL of AuNPs + 400 μL of CD21 at 20 ng·mL –1 + 100 μL of 1% BSA; and (d) 2.1 mL of AuNPs + 400 μL of CD21 at 20 ng·mL –1 + 100 μL of 1% BSA + 50 ng·mL –1 of gp350.

Article Snippet: Recombinant human CD21 and recombinant EBV gp350 proteins were purchased from Sino Biological Inc. All reagents and chemicals employed in this study were analytical grade and used without further purification.

Techniques:

Journal: Analytical Chemistry

Article Title: Early Detection of Multiple Sclerosis Using Spectroscopic Epstein–Barr Virus Biosensors Based on Infection Mechanism Mimicry

doi: 10.1021/acs.analchem.5c03964

Figure Lengend Snippet: (A) UV spectra and (B) bar chart that belong to the optimization of the CD21–gp350 interaction incubation temperature. (a) BSA@CD21@AuNP, (b) BSA@CD21@AuNP + 50 ng·mL –1 gp350 for 4 °C, (c) 25 °C, (d) 37 °C, and (e) 45 °C incubation temperature.

Article Snippet: Recombinant human CD21 and recombinant EBV gp350 proteins were purchased from Sino Biological Inc. All reagents and chemicals employed in this study were analytical grade and used without further purification.

Techniques: Incubation

Journal: Analytical Chemistry

Article Title: Early Detection of Multiple Sclerosis Using Spectroscopic Epstein–Barr Virus Biosensors Based on Infection Mechanism Mimicry

doi: 10.1021/acs.analchem.5c03964

Figure Lengend Snippet: (A) UV spectra and (B) Excel plot that belong to the optimization of the CD21–gp350 interaction incubation time. (a) BSA@CD21@AuNP, (b) BSA@CD21@AuNP + 50 ng·mL –1 gp350 for 15 min, (c) 30 min, (d) 60 min, and (e) 90 min incubation time.

Article Snippet: Recombinant human CD21 and recombinant EBV gp350 proteins were purchased from Sino Biological Inc. All reagents and chemicals employed in this study were analytical grade and used without further purification.

Techniques: Incubation

Journal: Analytical Chemistry

Article Title: Early Detection of Multiple Sclerosis Using Spectroscopic Epstein–Barr Virus Biosensors Based on Infection Mechanism Mimicry

doi: 10.1021/acs.analchem.5c03964

Figure Lengend Snippet: Analytical characteristics of the developed colorimetric EBV biosensors. (A) UV spectra of (a) BSA@CD21@AuNP, (b) 10 ng·mL –1 , (c) 25 ng·mL –1 , (d) 50 ng·mL –1 , (e) 80 ng·mL –1 , and (f) 100 ng·mL –1 gp350 additions. (B) Calibration plot of increasing concentrations of gp350 from 10 ng·mL –1 to 100 ng·mL –1 .

Article Snippet: Recombinant human CD21 and recombinant EBV gp350 proteins were purchased from Sino Biological Inc. All reagents and chemicals employed in this study were analytical grade and used without further purification.

Techniques:

Journal: Analytical Chemistry

Article Title: Early Detection of Multiple Sclerosis Using Spectroscopic Epstein–Barr Virus Biosensors Based on Infection Mechanism Mimicry

doi: 10.1021/acs.analchem.5c03964

Figure Lengend Snippet: UV spectra demonstrating the absorbance difference of a 1:1 ratio of (a) BSA@CD21@AuNP, (b) gp350 @BSA@CD21@AuNP, and (c) A27L + KMP11 + gp350 @BSA@CD21@AuNP.

Article Snippet: Recombinant human CD21 and recombinant EBV gp350 proteins were purchased from Sino Biological Inc. All reagents and chemicals employed in this study were analytical grade and used without further purification.

Techniques:

Journal: Nature Immunology

Article Title: Somatic deficiency of the human E3 ubiquitin ligase CBL in leukocytes impairs B cell but not T cell development and function

doi: 10.1038/s41590-025-02381-7

Figure Lengend Snippet: a , Counts of peripheral B cells in individuals with CBL -LOH over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. b , IgG levels in individuals with CBL -LOH (P1–P8) over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. c , Quantification of the indicated B cell subsets in the peripheral cryopreserved mononuclear cell blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2), heterozygous individuals ( n = 3). Data are shown as mean ± s.d. The statistical significance of differences was assessed in multiple two-sided Mann–Whitney tests, with correction for multiple testing; ** P < 0.005 and *** P < 0.0005. y.o., years old. d , Frequency of B cell subsets in cryopreserved PBMCs from HDs and individuals with CBL -LOH as determined by flow cytometry; HDs ( n = 13), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed using multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. e , CD5, CD9, CD21 and CD38 expression on transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 19), individuals with CBL -LOH ( n = 6). Data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05, ** P < 0.005 and **** P < 0.00005. f , Percentage of CD21 hi cells among transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 9), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed by Mann–Whitney test; *** P < 0.0005; gMFI, geometric mean fluorescence intensity.

Article Snippet: 162Dy , CD21 , REA940 , 130-124-315 , Miltenyi Biotec.

Techniques: Mass Cytometry, MANN-WHITNEY, Flow Cytometry, Expressing, Fluorescence

Journal: Nature Immunology

Article Title: Somatic deficiency of the human E3 ubiquitin ligase CBL in leukocytes impairs B cell but not T cell development and function

doi: 10.1038/s41590-025-02381-7

Figure Lengend Snippet: ( a ) Quantification of B-cell subsets in cryopreserved PBMCs from healthy donors, heterozygous carriers, and CBL-LOH patients of the indicated ages by mass cytometry. Controls 0–3 y.o. (n = 2); controls 4–15 y.o. (n = 9); controls 16–100 y.o. (n = 28); pediatric CBL-LOH (n = 5); adult CBL-LOH (n = 2); heterozygous individuals (n = 3). Mean ± s.d. Statistical significance was assessed using two-sided Mann–Whitney tests with correction for multiple testing. ( b ) Frequency of B-cell subsets in healthy donors (n = 10) and CBL-LOH patients (n = 4) by flow cytometry. Mean ± s.d. Significance was evaluated with two-sided Mann–Whitney tests adjusted for multiple comparisons. **p < 0.005. ( c ) MFI of B-cell markers on CD21 lo and transitional B cells from healthy donors (n = 6) and patients P1–3 (n = 3). Lines show means. ( d–f ) Modeling the CBL ΔExon8 variant in primary human CD34⁺ HSPCs. ( d ) Agarose gel electrophoresis of PCR products from AAVS1 or CBL loci 72 h after nucleofection; representative of >5 biological replicates. ( e ) Editing efficiency using sgRNA pair 1+2 by NGS, showing ~80% of the ~400 bp exon-8 deletion. ( f ) Western blot of CBL protein levels after CBL or AAVS1 editing. ( g ) Quantification of three biological replicates from (f). Mean ± s.d. ( h ) NGS quantification of exon-8 deletions for all three guide pairs (n = 2 biological replicates). Bars show means. ( i ) Editing efficiencies at days 8 and 21 in differentiation cultures by NGS. Mean ± s.d. from three biological replicates. ( j–m ) Bulk RNA-seq of AAVS1- and CBL-edited CD19⁺CD10⁺CD20 low HSPC-derived B-cell progenitors. ( j ) Gene-set enrichment analysis showing significantly enriched (red) or depleted (blue) pathways (NES: normalized enrichment score). ( k,l ) Differential expression of leading-edge genes in the ( k ) Hallmark G2–M checkpoint and ( l ) Hallmark mTORC1 signaling gene sets (Z-transformed normalized counts). ( m ) Transcriptional overlap between CBL-edited and PI3K GOF progenitors; Venn diagrams show shared significantly up- or downregulated genes, with overlap significance by binomial test.

Article Snippet: 162Dy , CD21 , REA940 , 130-124-315 , Miltenyi Biotec.

Techniques: Mass Cytometry, MANN-WHITNEY, Flow Cytometry, Variant Assay, Agarose Gel Electrophoresis, Western Blot, RNA Sequencing, Derivative Assay, Quantitative Proteomics, Transformation Assay

Journal: Frontiers in Immunology

Article Title: Characteristics of tertiary lymphoid structures in prostate cancer and the impact of neoadjuvant therapy on their formation and maturation

doi: 10.3389/fimmu.2025.1663396

Figure Lengend Snippet: Clinicopathological features and prognostic significance of tertiary lymphoid structures (TLS) (A) The distribution of clinical features and immature density for the 27 patients in cohort 1. (B) Kaplan-Meier survival curves for progression-free survival (PFS) of 27 patients with no treatment before surgery stratified based on the median density of TLSs (B) , density of intra-/peri- TLSs (C) , maturation of TLSs (D) and the existence of SFL-TLSs (In SFL-TLS, CD21+ follicular dendritic cells form dense clusters within the core germinal center) (E) . B–E: Differentiate Kaplan-Meier curves with distinct colors for better visual clarity. 3D & 3E: Clarify the difference between “mature TLS” and “SFL-TLS.”.

Article Snippet: Then, slides were treated by microwave to induce antigen retrieval using citric acid solution for 15 min. 6 primary antibodies were used, including CD4 (1:500, #Ab67480, Abcam), CD8 (1:300, #66868-1-Ig, proteintech), CD20 (1:1000, #60271-1-Ig, proteintech), CD21 (1:200, #24374-1-AP, proteintech), Ki67(1:1000, #HA721115, HuaBio), pan-cytokeratin (1:2000, # HA601094 , HuaBio), DAPI (1:500, #C1002, Beyotime).

Techniques:

Journal: Frontiers in Immunology

Article Title: Characteristics of tertiary lymphoid structures in prostate cancer and the impact of neoadjuvant therapy on their formation and maturation

doi: 10.3389/fimmu.2025.1663396

Figure Lengend Snippet: Characteristics of tertiary lymphoid structures (TLS) in treatment-naive group and neoadjuvant hormone therapy (NHT) group in cohort 2. (A) mIHC of immature and mature TLS in NHT group and treatment-naive group in cohort 2; (B) Comparison of the ratio of CD4 + cells, CD8+ cells, CD20+cells and CD21+cells in tumor microenvironment (TME) in the NHT group and treatment-naive group. (C) Proportion of different TLSs maturation stage in the NHT group and treatment-naive group. (D) Proportion of different density of TLS in the NHT group and treatment-naive group. (E) Proportion of different location of TLS in the NHT group and treatment-naive group. P values are denoted as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ***p < 0.0001.

Article Snippet: Then, slides were treated by microwave to induce antigen retrieval using citric acid solution for 15 min. 6 primary antibodies were used, including CD4 (1:500, #Ab67480, Abcam), CD8 (1:300, #66868-1-Ig, proteintech), CD20 (1:1000, #60271-1-Ig, proteintech), CD21 (1:200, #24374-1-AP, proteintech), Ki67(1:1000, #HA721115, HuaBio), pan-cytokeratin (1:2000, # HA601094 , HuaBio), DAPI (1:500, #C1002, Beyotime).

Techniques: Comparison

Journal: Frontiers in Immunology

Article Title: Characteristics of tertiary lymphoid structures in prostate cancer and the impact of neoadjuvant therapy on their formation and maturation

doi: 10.3389/fimmu.2025.1663396

Figure Lengend Snippet: Characteristics and changes of immune cells in pre- and post- neoadjuvant hormone therapy (NHT) groups in GSE111177 . (A) Supervised clustering of PCa specimens by NHT (n = 24), displaying ssGSEA scores ( GSE111177 ). (B) The biological process of DEGs identified in pre-/post- NHT groups ( GSE111177 ). (C) Comparison of the expression of CD4, CD8A and CD8B, CD20, FOXP3, CD21, GZMB, PRF1 in PCa patients before and after NHT using GSE111177 cohort (n = 24). (D) Comparison of GSVA scores utilizing four TLS signatures in 24 paired pre-NHT and post-NHT samples from GSE111177 .

Article Snippet: Then, slides were treated by microwave to induce antigen retrieval using citric acid solution for 15 min. 6 primary antibodies were used, including CD4 (1:500, #Ab67480, Abcam), CD8 (1:300, #66868-1-Ig, proteintech), CD20 (1:1000, #60271-1-Ig, proteintech), CD21 (1:200, #24374-1-AP, proteintech), Ki67(1:1000, #HA721115, HuaBio), pan-cytokeratin (1:2000, # HA601094 , HuaBio), DAPI (1:500, #C1002, Beyotime).

Techniques: Comparison, Expressing

Journal: Frontiers in Immunology

Article Title: Characteristics of tertiary lymphoid structures in prostate cancer and the impact of neoadjuvant therapy on their formation and maturation

doi: 10.3389/fimmu.2025.1663396

Figure Lengend Snippet: Characterization of tumor immune microenvironment (TME) and tertiary lymphoid structures (TLS) before and after neoadjuvant hormone therapy (NHT). (A) Comparison of the ratio of CD4 + cells, CD8+ cells, CD20+cells and CD21+cells in TME before and after NHT. (B) Comparison of the TLS presence in TME before and after NHT. (C) Comparison of the intra-tumor TLS presence in TME before and after NHT. (D) Comparison of the TLS maturation stage in TME before and after NHT. (E) Evaluation of TLS maturation stage using mIHC staining of CD4, CD8 CD20, CD21 before and after NHT. P values are denoted as follows: **p < 0.01, ***p < 0.001, NS p>0.05.

Article Snippet: Then, slides were treated by microwave to induce antigen retrieval using citric acid solution for 15 min. 6 primary antibodies were used, including CD4 (1:500, #Ab67480, Abcam), CD8 (1:300, #66868-1-Ig, proteintech), CD20 (1:1000, #60271-1-Ig, proteintech), CD21 (1:200, #24374-1-AP, proteintech), Ki67(1:1000, #HA721115, HuaBio), pan-cytokeratin (1:2000, # HA601094 , HuaBio), DAPI (1:500, #C1002, Beyotime).

Techniques: Comparison, Staining