hrp conjugated mouse anti human β actin antibody  (Proteintech)

Journal: Frontiers in Immunology

Article Title: High interleukin-35 expression is associated with the severity of rheumatic mitral stenosis

doi: 10.3389/fimmu.2025.1537497

Figure Lengend Snippet: IL-35 expression in MV tissues of RMS patients and controls. (A) Western blot analysis of IL-35 expression in controls, moderate and severe RMS patients. β-actin was used as an internal control. (B) IL-35 expression in the MV tissues of RMS patients compared to controls (MVP). (C) Comparisons of IL-35 expressions in MV tissues of moderate and severe RMS patients vs. controls (MVP). (D) A positive correlation between p35 expression in MV tissue and the severity of RMS. ns, non-significant; **p < 0.01.

Article Snippet: As an internal control, mouse anti-human β-actin mAb (Proteintech, USA) and peroxidase-conjugated anti-mouse IgG (H+L) (Proteintech, USA) were used.

Techniques: Expressing, Western Blot, Control

Journal: Nature Communications

Article Title: MANF stimulates autophagy and restores mitochondrial homeostasis to treat autosomal dominant tubulointerstitial kidney disease in mice

doi: 10.1038/s41467-023-42154-0

Figure Lengend Snippet: a RNA-seq of WT and mutant TALs at 16 weeks. n = 5/group. GO biological process perturbation bar plot showed the most upregulated pathways in the mutant vs. WT TALs at 16 weeks. GAGE analysis. b Quantitative PCR for proinflammatory cytokines, normalized to β-actin, in isolated TALs (WT: n = 6 for Icam1 and n = 8 for other genes; Umod DEL/+ : n = 4 for Tnfa , Il6 and Ccl2 , n = 7 for Il1b and n = 5 for Icam1 ) at 16 weeks. Two-tailed t -test, Mean ± SD. p = 0.0039 (for Tnfa ), 0.0075 ( Il1b ), 0.0179 ( Il6 ), 0.0151 ( Icam1 ), 0.0002 ( Ccl2 ). c H&E staining of paraffin kidney sections from Umod +/+ and Umod DEL/+ mice at 24 weeks. Red arrow indicates interstitial inflammatory infiltration. Black arrow indicates a cortical renal cyst. d , e Representative IF staining of macrophage (F4/80) and UMOD with nuclei counterstain (blue) on frozen kidney sections from Umod +/+ and Umod DEL/+ mice at 24 weeks ( d ) with quantification ( e ). n = 10 images/genotype. Two-tailed t -test, Mean ± SD. **** p < 0.0001. f Immunoblot analysis monitoring STING signaling in Umod +/+ and Umod DEL/+ TALs at 16 weeks, including STING, p-TBK1/TBK1, p-NF-κB p65/NF-κB p65 (WT: n = 4; Umod DEL/+ : n = 3) and p-IRF3/IRF3 (WT: n = 3; Umod DEL/+ : n = 4) with densitometry analysis. Two-tailed t -test, Mean ± SD. p = 0.0008 (for p-TBK1/TBK1), 0.0493 (p-NF-κB/NF-κB), 0.0484 (p-IRF3/IRF3). g Immunoblot analysis monitoring cleaved CASP9 and CASP3 in Umod +/+ ( n = 4) and Umod DEL/+ ( n = 6) kid n eys at 16 weeks with densitometry analysis. Two-tailed t -test, Mean ± SD. p = 0.0176 (for cleaved CASP9), 0.0012 (cleaved CASP3). h Representative IF staining of TUNEL and UMOD (red) with nuclei counterstain (blue) on frozen kidney sections from Umod +/+ and Umod DEL/+ mice at 24 weeks. The TUNEL-positive cells were quantified. n = 5 images/genotype. Red and white arrows: UMOD - and UMOD + cells, respectively. Two-tailed t -test, Mean ± SD. **** p < 0.0001. Source data are provided as a Source Data file.

Article Snippet: To ensure equal protein loading, the same blot was stripped with stripping buffer (25 mM glycine + 1% SDS, pH = 2.0) and then incubated with a HRP-conjugated anti-mouse (human) β-actin antibody (Sigma-Aldrich) or anti-mouse (human) GAPDH antibody (Cell Signaling, Danvers, MA).

Techniques: RNA Sequencing Assay, Mutagenesis, Real-time Polymerase Chain Reaction, Isolation, Two Tailed Test, Staining, Western Blot, TUNEL Assay