rabbit polyclonal anti gapdh  (Proteintech)

Journal: mBio

Article Title: A bacterial family of fatty acid acyltransferases related to the Shigella effector IcsB

doi: 10.1128/mbio.03890-25

Figure Lengend Snippet: A subset of novel k-FATs exhibits yeast toxicity and IcsB-like fatty acid acyltransferase activity. ( A ) Map of the pRS313 Gal1p2 centromeric plasmid used for the galactose-inducible expression of IcsB and its homologs. ( B )The expression of IcsB and seven of its homologs in the presence of galactose inhibits yeast growth compared to the empty vector (EV) control. The glucose plate indicates that there is no growth inhibition in the absence of expression and that similar amounts of each strain were plated. Four dilutions were plated (non-diluted, 1/10, 1/100, and 1/1,000). The graph represents the quantification of growth on three independent replicates of the 1/1,000 dilution. The error bars represent the standard deviation of the mean. A Student’s t-test with unpaired data and a 95% CI was performed (*, P < 0.05; **, P < 0.1, ***, P < 0.01). ( C ) Protein acylation in the membrane protein fraction of yeast expressing IcsB and its homologs according to the in-gel fluorescence acyltransferase assay (upper panels). -Alk IcsB is the IcsB sample prior to the addition of Alk-16. -Alk IcsB, Alk-16 EV, and Alk-16 IcsB were loaded on both gels to serve as controls. This acylation assay was performed using the strains from B that were induced with galactose for six hours in the presence of Alk-16 for the final 4 h. The total protein in these samples were assessed using the TGX stain (bottom panels). ( D ) The expression of IcsB and of its homologs was assessed by immunoblotting with an antibody binding to their 3× FLAG tag and with the GAPDH as a loading control.

Article Snippet: Primary antibodies used were mouse FLAG-tag (Sigma-Aldrich, F3165), mouse Myc-tag (Genescript, A00704), and rabbit GAPDH (Bioss antibodies, BS-8789R) diluted 1/5,000, 1/1,000, and 1/10,000, respectively.

Techniques: Activity Assay, Plasmid Preparation, Expressing, Control, Inhibition, Standard Deviation, Membrane, Fluorescence, Staining, Western Blot, Binding Assay, FLAG-tag

Journal: mBio

Article Title: A bacterial family of fatty acid acyltransferases related to the Shigella effector IcsB

doi: 10.1128/mbio.03890-25

Figure Lengend Snippet: Mutations in the catalytic site of k-FATs abolish their enzymatic activity. ( A ) Yeast growth inhibition tests of catalytic residues mutants H145A (left), D195A (center), and C306A (right) compared to their WT counterpart in IcsB and in toxic homologs. The graph represents the quantification of growth on three independent replicates with the dilution of yeast cultures 1/1,000. The error bars represent the standard deviation from the mean. A Student’s t -test with unpaired data and a 95% CI was performed (*, P < 0.05; **, P < 0.1, ***, P < 0.01). ( B ) In-gel protein acylation of C306A mutants compared to their WT counterparts (upper panels). The total protein in these samples were assessed using the TGX stain (bottom panels). ( C ) The expression of IcsB and toxic homolog C306A mutants was compared to their WT counterparts by immunoblotting with an antibody binding to their 3×FLAG tag and with the GAPDH as a loading control.

Article Snippet: Primary antibodies used were mouse FLAG-tag (Sigma-Aldrich, F3165), mouse Myc-tag (Genescript, A00704), and rabbit GAPDH (Bioss antibodies, BS-8789R) diluted 1/5,000, 1/1,000, and 1/10,000, respectively.

Techniques: Activity Assay, Inhibition, Standard Deviation, Staining, Expressing, Western Blot, Binding Assay, Control

Journal: Biomedicines

Article Title: Ionic Extracts of Magnesium Powders Promote In Vitro Lymphangiogenesis

doi: 10.3390/biomedicines14040913

Figure Lengend Snippet: Effects of magnesium extracts on lymphangiogenesis-related gene and protein expression in LECs. ( A ) Relative Vegfa mRNA expression levels in LECs treated with Control, Mg (1:10), Mg (1:100), and Mg (1:1000), as determined by qRT-PCR. ( B ) Relative Vegfc mRNA expression levels in LECs under the same treatment conditions. ( C ) Representative Western blot images showing VEGFA and VEGFC protein expression in LECs after treatment with different concentrations of magnesium extracts, with GAPDH used as the internal loading control. ( D ) Additional Western blot analysis showing VEGFR3 protein expression under the same treatment conditions, with β-actin used as the internal loading control. Data in ( A , B ) are presented as the mean ± SD ( n = 3). Statistical analysis for ( A , B ) was performed using one-way ANOVA followed by Tukey’s multiple-comparisons test. Statistical significance is indicated as * p < 0.05, ** p < 0.01; ns, not significant.

Article Snippet: The primary antibodies used in this study included anti-VEGFA rabbit polyclonal antibody (ZEN-BIOSCIENCE Co., Ltd., Chengdu, China), anti-VEGFC rabbit polyclonal antibody (ZEN-BIOSCIENCE Co., Ltd., Chengdu, China), anti-VEGFR3 rabbit polyclonal antibody (Affinity Biosciences, Shanghai, China), anti-GAPDH rabbit polyclonal antibody (Affinity Biosciences, Shanghai, China), and anti-β-actin rabbit polyclonal antibody (Affinity Biosciences, Shanghai, China).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Structural insights into specific recognition of the histone variant H2A.Z by the YL1 subunit of SRCAP chromatin-remodeling complex

doi: 10.1016/j.jbc.2026.111294

Figure Lengend Snippet: Interactions between YL1-Z and H2A.Z–H2B in vivo . A , a, e: HEK 293T cells were transfected with GFP alone expressing plasmid for 24 h and imaged using fluorescence microscopy (100× magnification); b, f: HEK 293T cells were transfected with WT YL1 1–105 -GFP (YL1-GFP) expressing plasmid for 24 h and imaged using fluorescence microscopy (100× magnification); c, g: HEK 293T cells were transfected with YL1 1–105 F29A/Y30A/Y34A/F37A -GFP (YL1 4A -GFP) expressing plasmid for 24 h and imaged using fluorescence microscopy (100× magnification); d, h: HEK 293T cells were transfected with YL1 1–105 delete amino acids 29–37 -GFP (YL1 del -GFP) expressing plasmid for 24 h and imaged using fluorescence microscopy (100× magnification). B , interaction analysis between WT and mutant YL1 with H2A.Z–H2B. HEK 293T cells expressing WT YL1 and mutants were analyzed by Western blotting using primary antibodies against GFP and His tag, respectively. Compared with WT YL1, the binding capacity of mutants with H2A.Z–H2B was severely compromised. C , expression level analysis of GFP fusion proteins in HEK 293T cells. Expression levels of GFP, YL1-GFP, YL1 4A -GFP, and YL1 del -GFP in HEK 293T cells were analyzed using an antibody against GAPDH. The results show that GFP, YL1-GFP, YL1 4A -GFP, and YL1 del -GFP are expressed at comparable levels in HEK 293T cells. HEK, human embryonic kidney cell line.

Article Snippet: Western blot was performed according to the manufacturer's protocol (Sangon Biotech Co, Ltd, Anti-GFP mouse monoclonal antibody [order no.: D191040]; Anti-6X His Tag mouse monoclonal antibody [order no.: D191001]; Anti-GAPDH rabbit polyclonal antibody [order no.: D110016]; horseradish peroxidase–conjugated Goat Anti-Mouse IgG [order no.: D110087]; and horseradish peroxidase–conjugated Goat Anti-Rabbit IgG [order no.: D110058]).

Techniques: In Vivo, Transfection, Expressing, Plasmid Preparation, Fluorescence, Microscopy, Mutagenesis, Western Blot, Binding Assay