peroxidase goat polyclonal anti mouse  (Vector Laboratories)

Journal: Cell Reports Medicine

Article Title: FLT3L-based drug conjugate effectively targets chemoresistant leukemia stem cells in acute myeloid leukemia

doi: 10.1016/j.xcrm.2025.102365

Figure Lengend Snippet: Constructed FL-Fc-DM1 retains ligand-dependent signaling activity and selectively inhibits AML cells (A) Expression of rhFL-Fc and purification of rhFL-Fc via protein-A sepharose. Lane 1: pre-stained protein marker; lane 2: cell culture supernatant from HEK293 cells transfected with pFUSE-FL-hIgG4-Fc; lane 3: purified rhFL-Fc via protein-A sepharose column. (B) Immunoblotting analysis of final purified rhFL-Fc. Lane 1: pre-stained protein marker; lane 2: final purified rhFL-Fc; lane 3: immunoblotting of final purified rhFL-Fc using anti-FL polyclonal antibody. lane 4: pre-stained protein marker. (C) SDS-PAGE analysis of FL-Fc, FL-Fc-SMCC, and FL-Fc-DM1 under reducing and non-reducing conditions. Lanes 1 and 4: rhFL-Fc; lanes 2 and 5: FL-Fc-SMCC; lanes 3 and 6: FL-Fc-DM1. (D) UPLC analysis of FL-Fc-DM1 using an ACQUITY UPLC Protein BEH C4 Column. (E) Immunoblotting analysis of p-FLT3, FLT3, p-ERK1/2, ERK1/2, p-AKT, AKT, GAPDH in pre-serum-starved AML cells treated with FL-Fc-DM1 (100 ng/mL) or FL-Fc (100 ng/mL) for the indicated time durations. (F) Inhibition curves of FL-Fc-DM1, DM1, and FL-Fc on HCD-57 cells and HCD-57-derived cells stably expressing human wild-type FLT3, human FLT3-ITD mutant, or mouse wild-type FLT3. Data represent the mean ± SD. n = 3 biological replicates for each group. (G) Binding affinity measurement rhFLT3 ecto /rmFLT3 ecto to rhFL. See also D. (H) Cell cycle analysis and statistical analysis of AML cells treated by different drugs for 24 h. The cell cycle analysis is conducted using FlowJo. Data represent the mean ± SD. n = 3 biological replicates for each group. One-way ANOVA (Tukey’s post hoc test): ns, not significant; ∗ p < 0.05 and ∗∗ p < 0.01. (I) Immunoblotting analysis and statistical analysis of p-RB, and RB in MV-4-11 and THP-1 cells treated by FL-Fc-DM1 for the indicated time durations. Cells were treated with FL-Fc-DM1 (100 ng/mL for THP-1 and 50 ng/mL for MV-4-11). RB protein levels were analyzed by ImageJ. Data represent the mean ± SD. n = 3 biological replicates for each group. One-way ANOVA (Tukey’s post hoc test): ns, not significant; ∗ p < 0.05 and ∗∗ p < 0.01. (J) FACS and statistical analysis of the FITC + population in AML cells stably expressing Clover-hGEMININ (1-100) . Cells were treated with indicated concentration of drugs for 24 h. Data represent the mean ± SD. n = 3 biological replicates for each group. One-way ANOVA (Tukey’s post hoc test): ns, not significant; ∗ p < 0.05 and ∗∗ p < 0.01. (K) Results and statistical analysis of colony-forming numbers of AML cells treated with indicated concentrations of different drugs FL-Fc, DM1, FL-Fc + DM1, or FL-Fc-DM1 for 10 days. Data represent the mean ± SD. n = 3 biological replicates for each group. One-way ANOVA (Tukey’s post hoc test): ns, not significant; ∗ p < 0.05 and ∗∗ p < 0.01. (L) Schematic drawing of recombinant human FL-Fc-DM1. The upper blue region represents the FL moiety.

Article Snippet: Horse polyclonal secondary anti-mouse IgG, HRP-linked , Cell Signaling Technology , Cat#7076; RRID: AB_330924.

Techniques: Construct, Activity Assay, Expressing, Purification, Staining, Marker, Cell Culture, Transfection, Western Blot, SDS Page, Inhibition, Derivative Assay, Stable Transfection, Mutagenesis, Binding Assay, Cell Cycle Assay, Concentration Assay, Recombinant

Journal: Infection and Immunity

Article Title: Parenteral vaccination with recombinant EtpA glycoprotein impairs enterotoxigenic E. coli colonization

doi: 10.1128/iai.00601-24

Figure Lengend Snippet: Vaccination administration schedule impacts antibody diversity. ( A ) Schematic of EtpA lectin activity wherein the C-terminal repeat region of EtpA (CTR) directs binding to A blood group glycans expressed on the surface of enterocytes. ( B ) Negative stain electron microscopy polyclonal antibody epitope mapping (nsEMPEM) protocol. Created in BioRender. Fleckenstein, J. (2025) https://BioRender.com/ee1smdr ( C ) nsEMPEM of IgG Fabs generated from sera of mice vaccinated every two weeks or every three weeks with rEtpA adjuvanted with dmLT. Selected two-dimensional class averages of rEtpA in complex with Fabs generated from sera of mice vaccinated every two weeks (top row). The middle panel shows multiple-Fabs/rEtpA molecules, and the bottom row shows single-Fabs/rEtpA from mice vaccinated every three weeks with rEtpA/dmLT. Pseudo-coloring reflects rEtpA (blue), CTR-binding Fabs (orange), and NTD-binding Fabs (green).

Article Snippet: Anti-mouse IgG , Polyclonal horse anti-mouse IgG (H&L) affinity-purified , Cell Signaling , 7076 , AB_330924.

Techniques: Activity Assay, Binding Assay, Staining, Electron Microscopy, Generated

Journal: iScience

Article Title: A bacterial genotoxin reveals a p53-proteasome-LC3 regulatory axis that drives the suppression of autophagy in cells experiencing sublethal DNA damage

doi: 10.1016/j.isci.2025.112118

Figure Lengend Snippet:

Article Snippet: Horse polyclonal α-Mouse IgG, HRP-linked , Cell Signaling Technology, USA , Cat#7076; RRID: AB_330924.

Techniques: Virus, Recombinant, Cell Recovery, Modification, Gentle, Flow Cytometry, Reverse Transcription, Bicinchoninic Acid Protein Assay, RNA Extraction, Western Blot, Cloning, Mutagenesis, Plasmid Preparation, Software, Cell Culture, Microscopy, Nucleic Acid Electrophoresis