ra3 6b2  (Bio-Rad)

Journal: MethodsX

Article Title: Feline leukocyte immunophenotyping: an optimised whole-blood flow cytometry protocol

doi: 10.1016/j.mex.2026.103869

Figure Lengend Snippet: Representative leukocyte-gated histograms illustrating antibody titration for feline leukocyte immunophenotyping. All histograms display singlet leukocytes, defined by FSC-A versus SSC-A morphological gating, followed by FSC—H versus FSC-A singlet discrimination. Titration was performed for CD18, CD21, CD45R, CD4, CD5 and CD8 monoclonal antibodies using three antibody volumes: 10 µL (1:10 dilution), 5.0 µL (1:20 dilution), 3.0 µL (1:33 dilution) and 1.5 µL (1:66 dilution). Minimal working volumes were selected based on optimal signal-to-noise ratios.

Article Snippet: Step 2 – Leukocytes extracellular staining Materials • EDTA Whole blood sample ± Transfix® • 200 μl pipettes • 100 μl pipettes • 10 μl pipettes • Flow cytometry tubes • Permanent marker • Cytometer tube rack Reagents • Monoclonal antibodies: ○ CD5 Anti-cat – clone FE1.1B11 (BIO-RAD®) ○ CD4 Anti-cat – clone vpg34 (BIO-RAD®) ○ CD8 Anti-cat alpha/beta purified – clone vpg9 (BIO-RAD®) ○ Rat Anti-Mouse IgG1 – clone X56 (BIO-RAD®) ○ CD18 Mouse Anti-Dog – clone CA1.4E9 (BIO-RAD®) ○ CD21 Mouse Anti-Dog – clone CA2.1D6 (BIO-RAD®) ○ CD45R Rat Anti-Mouse – clone RA3–6B2 (BIO-RAD®) • 10x Red blood cells (RBC) lysis buffer solution (BD FACSTM lysing solution) • PBS 1% solution Equipment • Countess TM 3 (Thermo Fisher Scientific, USA) • Freezer • Dark incubation chamber • Timer • Vortex (MX-S®, China) • Centrifuge (model 5810R, Eppendorf®, Germany) • Flow cytometry analyser BD FACSCanto II (Becton Dickinson (BD), San Jose, USA) Methods 1.

Techniques: Titration, Bioprocessing

Journal: FASEB BioAdvances

Article Title: The Impact of Heterotopic Spleen Regeneration on Tumor Growth

doi: 10.1096/fba.2025-00254

Figure Lengend Snippet: Analysis of cell population in tumor of intact, SE and SE+ST animals on Days 7 (A), 12 (B) and 15 (C). Blue arrows—gating from CD3+ cell population, green arrows—gating from CD3− cell population. D, Percentage content of CD3+, CD8a+, CD4+, CD45R+, F4/80+Ly6C−, F4/80−Ly6C+, F4/80+Ly6C+ cells. (A–D) N = 3 per group. Statistical analysis was carried out by non‐parametric Kruskal–Wallis test with post hoc Dunn's test (mean with SEM), * р ‐value ≤ 0.05, ** p ‐value ≤ 0.005, *** p ‐value ≤ 0.0005.

Article Snippet: To determine the phenotypes of tumor‐infiltrating cells, isolated cells were stained for 1 h at room temperature in the dark with the following panel of anti‐mouse antibodies: CD8a antibody anti‐mouse VioBlue (130‐123‐865, Miltenyi Biotec, Germany), CD3 antibody anti‐mouse FITC, REAfinity (130‐119‐758, Miltenyi Biotec, Germany), F4/80‐PE REAfinity (130‐102‐422, Miltenyi Biotec, Germany), rat anti‐mouse CD4 StarBright Blue 700 (SBB700) (MCA2691SBB700, Biorad, USA), CD45R antibody anti‐mouse PE‐Vio 770 (130‐102‐817, Miltenyi Biotec, Germany), Ly‐6C antibody anti‐mouse APC REAfinity (130‐111‐779, Miltenyi Biotec, Germany).

Techniques:

Journal: JFMS Open Reports

Article Title: Feline malignant lymphoma in an uncommon location as a differential diagnosis for neurological disease

doi: 10.1177/20551169241300815

Figure Lengend Snippet: List of the antibodies used and results of immunohistochemical staining

Article Snippet: CD45R , Rat, clone B220 (Ly 5) , Cedarlane , 1:1000 , +.

Techniques: Immunohistochemical staining

Journal: JFMS Open Reports

Article Title: Feline malignant lymphoma in an uncommon location as a differential diagnosis for neurological disease

doi: 10.1177/20551169241300815

Figure Lengend Snippet: (a) The carotid body is infiltrated with a high number of monomorphic round cells. (b) The majority of cells is immunopositive for CD45R

Article Snippet: CD45R , Rat, clone B220 (Ly 5) , Cedarlane , 1:1000 , +.

Techniques: