rabbit anti pgk1  (Affinity Biosciences)

Journal: Antioxidants

Article Title: Boeravinone A Alleviates Oxidative Stress and Inflammation in LPS-Induced Acute Kidney Injury by Targeting PGK1

doi: 10.3390/antiox15070900

Figure Lengend Snippet: PGK1 is identified as a potential molecular target of BA. ( A ) Silver-stained SDS-PAGE gel showing proteins pulled down by BA. ( B ) Volcano plot of proteins identified by mass spectrometry, with PGK1 highlighted as the candidate target. ( C ) CETSA showing the thermal stability of PGK1 in BA- and DMSO-treated cells ( n = 3). ( D ) DARTS assay showing the effect of BA on PGK1 stability against Pronase E digestion ( n = 3). Assay demonstrating the effect of BA on PGK1 protein stability against Pronase E digestion ( n = 3). ( E ) Coomassie Blue staining of purified PGK1 protein. ( F ) ITC analysis of the binding affinity between BA and PGK1. ( G ) Molecular docking model showing the predicted binding mode of BA with PGK1. Data are presented as the mean ± SD. n = 3 for CETSA and DARTS analyses.

Article Snippet: Anti-PGK1 (A12686) antibodies were obtained from ABclonal (Wuhan, China).

Techniques: Staining, SDS Page, Mass Spectrometry, Purification, Binding Assay

Journal: Antioxidants

Article Title: Boeravinone A Alleviates Oxidative Stress and Inflammation in LPS-Induced Acute Kidney Injury by Targeting PGK1

doi: 10.3390/antiox15070900

Figure Lengend Snippet: Virtual KO of PGK1 reveals its regulatory role in macrophage inflammatory responses. ( A ) Volcano plot showing significantly affected genes after PGK1 virtual KO in LPS-stimulated RAW264.7 macrophages. ( B ) Top genes significantly affected by PGK1 virtual KO, ranked by z-score. ( C ) GO BP enrichment analysis of genes affected by PGK1 virtual KO, showing enrichment in immune and inflammation-related biological processes.

Article Snippet: Anti-PGK1 (A12686) antibodies were obtained from ABclonal (Wuhan, China).

Techniques:

Journal: Antioxidants

Article Title: Boeravinone A Alleviates Oxidative Stress and Inflammation in LPS-Induced Acute Kidney Injury by Targeting PGK1

doi: 10.3390/antiox15070900

Figure Lengend Snippet: PGK1 is involved in BA-mediated activation of the Keap1-Nrf2 pathway and reduction in oxidative stress. ( A ) Western blot validation of Pgk1 knockdown efficiency. Cells were treated with LPS (1 μg/mL) and BA (20 μM) for 24 h. ( B ) Western blot analysis of Nrf2 protein expression in LPS-stimulated shNC and sh Pgk1 macrophages ( n = 3). ( C ) qRT-PCR analysis of Keap1 and Nfe2l2 mRNA expression ( n = 3). ( D ) qRT-PCR analysis of Nqo1 , Gclc , and Gclm mRNA expression ( n = 3). ( E ) Immunofluorescence staining showing Nrf2 nuclear translocation. Nrf2 is shown in red, nuclei are stained with DAPI (blue) ( F ) Representative ROS staining images in different treatment groups. ROS is shown in green (DCFH-DA) ( n = 3). Data is presented as the mean ± SD.

Article Snippet: Anti-PGK1 (A12686) antibodies were obtained from ABclonal (Wuhan, China).

Techniques: Activation Assay, Western Blot, Biomarker Discovery, Knockdown, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Translocation Assay

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Proteomic and Phosphoproteomic Characteristics of the Retina, Choroid, and Sclera in Guinea Pigs with Form-Deprivation Myopia

doi: 10.1016/j.mcpro.2025.101069

Figure Lengend Snippet: Expression analysis of characteristic proteins in retina, choroid, and sclera . A and B , quantification of NTRK2, Rho, and GNGT2 expression at the protein level by Western blotting in the retina of guinea pigs in NC and FDM groups (n = 3). C and D , quantification of SYK, BTK, and NF-κB p65 expression at the protein level by Western blotting in the choroid of guinea pigs in NC and FDM groups (n = 3). E and F , quantification of ACTN3, PGK1, TPM2, and TNNT3 expression at the protein level by Western blotting in the sclera of guinea pigs in the NC and FDM groups (n = 3). ACTIN was used as the loading control. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. FDM, form-deprivation myopia; NC, normal control.

Article Snippet: After incubating with the primary antibodies against NTRK2 (1:1000; ABclonal Technology), GNGT2 (1:1000; ABclonal Technology), Rho (1:1000; Affinity Biosciences), SYK (1:1000; ABclonal Technology), BTK (1:1000; ABclonal Technology), NF-κB p65 (1:1000; ABclonal Technology), ACTN3 (1:1000; ABclonal Technology), PGK1 (1:1000; ABclonal Technology), TPM2 (1:1000; ABclonal Technology), TNNT3 (1:1000; Affinity Biosciences), pan phospho-threonine (1:1000; ABclonal Technology), and SF3B1 (1:1000; SANTA) overnight at 4 °C, the polyvinylidene difluoride membranes were washed with TBST three times and then incubated with an HRP-conjugated secondary antibody at room temperature for 1 h. Images were captured using a chemiluminescence imaging system and quantified with AIWBwell (Servicebio).

Techniques: Expressing, Western Blot, Control