mouse anti ace primary antibody  (R&D Systems)

Journal: Signal Transduction and Targeted Therapy

Article Title: Bioengineered iPSC-derived human macrophages with increased angiotensin-converting enzyme (ACE) expression suppress solid tumor growth

doi: 10.1038/s41392-026-02650-3

Figure Lengend Snippet: Analysis of anti-tumor macrophage activation in the TME post-iMac treatment. Tumors were harvested and single-cell suspensions were prepared for flow cytometry analysis as detailed in the methods section (refer to Fig. ). a Gating strategy for distinguishing M1 and M2 macrophages using flow cytometry based on surface marker immunostaining. b Assessment of CD86 + (M1) and CD206 + (M2) expression in tumor associated macrophages (TAMs) in SK-MEL-28-derived xenografts (±Dox). The bar plot presents FACS quantification of M1 and M2 populations ( n = 5/group). c Assessment of arginase expression in TAMs. d ACE expression in TAMs. e – g Measurement of proinflammatory and anti-tumor markers IFN-γ, IL-12, and iNOS (left: representative histogram; right: mean fluorescence intensity) in TAMs in melanoma xenografts ( n = 5/group). Statistical analysis was performed using a two-sided unpaired Student’s t -test for comparisons in ( b , d , g ), and one-way ANOVA with Bonferroni’s correction for multiple comparisons in ( c , e , f ). Data are presented as means ± SEM. * p < 0.05 and **** p < 0.0001

Article Snippet: For ACE immunostaining, cells were first treated with a mouse anti-ACE primary antibody (R&D Systems, MAB9291, 1:100), followed by an Alexa Fluor 647 anti-mouse secondary antibody (Invitrogen, A-21235, 1:500).

Techniques: Activation Assay, Single Cell, Flow Cytometry, Marker, Immunostaining, Expressing, Derivative Assay, Fluorescence

Journal: PLOS One

Article Title: pET28g: A Golden Gate-compatible pET vector for protein expression in Escherichia coli , validated by production of functional human ACE2

doi: 10.1371/journal.pone.0327341

Figure Lengend Snippet: Protein expression as detected in total soluble protein extracts by western blot with anti-ACE2 antibody (A and B) and Coomassie Brilliant Blue staining (C and D) . A and C, soluble protein extracts of bacteria transformed with pET28a plasmid expressing ACE2 with a 6x-His tag at the N-terminus; B and D, soluble protein extracts of bacteria transformed with pET28g plasmids expressing ACE2 with an 8 × His-Gb1 tag (Gb1) or an MBP tag (MBP) at the N-terminus. NI – Soluble protein extract from bacteria before protein expression induction; I4 – Soluble protein extract from bacteria four hours after protein expression induction; M – Molecular weight marker (MB09002, NZYTech).

Article Snippet: Detection of ACE2 was performed using an anti-ACE2 primary antibody (1:2,000 dilution, AAF933 from R&D Systems) and HRP-conjugated anti-goat secondary antibody (1:20,000 dilution, sc-2020 from Santa Cruz Biotechnology).

Techniques: Expressing, Western Blot, Staining, Bacteria, Transformation Assay, Plasmid Preparation, Molecular Weight, Marker

Journal: PLOS One

Article Title: pET28g: A Golden Gate-compatible pET vector for protein expression in Escherichia coli , validated by production of functional human ACE2

doi: 10.1371/journal.pone.0327341

Figure Lengend Snippet: × His-Gb1 tag . A) SDS-PAGE analysis of fractions from the reverse HisTrap purification step, performed after TEV protease cleavage to remove the 8 × His-GB1 tag and His-tagged TEV protease. The flowthrough (FT) contains the cleaved (untagged) ACE2, while the elution fraction contains the His-tagged TEV protease and the cleaved His-tag. Washes 1 and 2 (W1, W2) with PBS pH 7.4 ensured complete recovery of untagged ACE2. The figure also shows fractions (a) and (b) collected from size-exclusion chromatography. (B) Size-exclusion chromatography profile of untagged ACE2 purified using a HiLoad 16/600 Superdex™ 200 pg column, where peak (b) corresponds to the untagged ACE2 protein. M – molecular weight marker (MB09002, NZYTech).

Article Snippet: Detection of ACE2 was performed using an anti-ACE2 primary antibody (1:2,000 dilution, AAF933 from R&D Systems) and HRP-conjugated anti-goat secondary antibody (1:20,000 dilution, sc-2020 from Santa Cruz Biotechnology).

Techniques: SDS Page, Purification, Size-exclusion Chromatography, Molecular Weight, Marker

Journal: PLOS One

Article Title: pET28g: A Golden Gate-compatible pET vector for protein expression in Escherichia coli , validated by production of functional human ACE2

doi: 10.1371/journal.pone.0327341

Figure Lengend Snippet: (A) Far-UV CD spectra of ACE2 with a characteristic α-helical profile, indicating a predominantly helical secondary structure. Measurements were performed at a protein concentration of 0.2 mg/mL in PBS, pH 7.4, using a 0.1 cm path length cuvette at 25°C. Baseline correction was applied using the buffer CD spectrum. (B) Rates of Mca-APK(Dnp) substrate cleavage by recombinant ACE2. The rates are given in fluorescence arbitrary units (a.u.) because the unquenched Mca substrate was not available to prepare a calibration curve. For comparison, the rates observed for the control reactions, in the absence of substrate or protein, were plotted. Differences between the rate determined for ACE2 and the controls are statistically significant.

Article Snippet: Detection of ACE2 was performed using an anti-ACE2 primary antibody (1:2,000 dilution, AAF933 from R&D Systems) and HRP-conjugated anti-goat secondary antibody (1:20,000 dilution, sc-2020 from Santa Cruz Biotechnology).

Techniques: Circular Dichroism, Protein Concentration, Recombinant, Fluorescence, Comparison, Control