fabp5 d1a7t rabbit mab  (Cell Signaling Technology Inc)

Journal: iScience

Article Title: OBP2A regulates epidermal barrier function and protects against cytotoxic small hydrophobic molecules

doi: 10.1016/j.isci.2024.111093

Figure Lengend Snippet:

Article Snippet: FABP5 (D1A7T) Rabbit mAb , Cell Signaling Technology , 39926.

Techniques: Imaging, Recombinant, Transfection, Saline, Sterility, Cell Culture, Protease Inhibitor, DC Protein Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Quantitation Assay, Fluorescence, Sequencing, Software

Journal: International Journal of Molecular Sciences

Article Title: Salivary and Lacrimal Gland Alterations of the Epidermal Fatty Acid-Binding Protein (E-FABP) in Non-Obese Diabetic Mice

doi: 10.3390/ijms23073491

Figure Lengend Snippet: Comparison of E-FABP concentrations in tears and saliva between the wild-type (WT) mice and the NOD mice. ( A ) Tear E-FABP concentration in the NOD mice was significantly lower than in the WT mice. ( p = 0.0135). ( B ) Saliva E-FABP concentration in the NOD mice was also significantly lower than in the WT mice. ( p = 0.04). * represents p < 0.05. E-FABP: epidermal fatty acid binding protein.

Article Snippet: The peroxidase system Vectastain ABC kit (rabbit IgG; Vector Laboratories, Burlingame, CA, USA) and the anti-FABP5 (D1A7T) rabbit monoclonal antibody (Cell Signaling Technology, Danvers, MA, USA) diluted with goat blocking serum at 1:200 were used.

Techniques: Concentration Assay, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Salivary and Lacrimal Gland Alterations of the Epidermal Fatty Acid-Binding Protein (E-FABP) in Non-Obese Diabetic Mice

doi: 10.3390/ijms23073491

Figure Lengend Snippet: Comparison of E-FABP immunohistochemistry in the salivary gland between the wild-type (WT) mice and the NOD mice. Representative immunohistochemistry images showed less intense immunostaining in the acinar epithelium of the NOD mice ( B ) when compared with the WT mice ( A ) (yellow arrows). IHC: immunohistochemistry.

Article Snippet: The peroxidase system Vectastain ABC kit (rabbit IgG; Vector Laboratories, Burlingame, CA, USA) and the anti-FABP5 (D1A7T) rabbit monoclonal antibody (Cell Signaling Technology, Danvers, MA, USA) diluted with goat blocking serum at 1:200 were used.

Techniques: Immunohistochemistry, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Salivary and Lacrimal Gland Alterations of the Epidermal Fatty Acid-Binding Protein (E-FABP) in Non-Obese Diabetic Mice

doi: 10.3390/ijms23073491

Figure Lengend Snippet: Comparison of E-FABP immunohistochemistry in the lacrimal glands of the wild-type (WT) mice and the NOD mice. Representative immunohistochemistry images showed increased immunostaining of the acinar epithelium in the NOD mice ( B , D ) compared to the WT mice ( A , C ) (yellow arrows). Moreover, a denser infiltration of inflammatory cells around the acinar units of the NOD mice compared to the WT mice can be observed (green arrow). IHC: immunohistochemistry.

Article Snippet: The peroxidase system Vectastain ABC kit (rabbit IgG; Vector Laboratories, Burlingame, CA, USA) and the anti-FABP5 (D1A7T) rabbit monoclonal antibody (Cell Signaling Technology, Danvers, MA, USA) diluted with goat blocking serum at 1:200 were used.

Techniques: Immunohistochemistry, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Salivary and Lacrimal Gland Alterations of the Epidermal Fatty Acid-Binding Protein (E-FABP) in Non-Obese Diabetic Mice

doi: 10.3390/ijms23073491

Figure Lengend Snippet: Comparison of E-FABP mRNA expressions in the salivary and lacrimal glands of the wild-type (WT) and the NOD mice. ( A ) In the salivary glands, the E-FABP mRNA expression in the NOD mice was significantly lower than that in the WT mice ( p = 0.001). ( B ) In the lacrimal glands, the E-FABP mRNA expression in the NOD mice was significantly higher than that in the WT mice ( p = 0.0008). * and ** represent p < 0.05 and p < 0.001, respectively. SG: salivary gland, LG: lacrimal gland, GAPDH: glyceraldehyde phosphate dehydrogenase.

Article Snippet: The peroxidase system Vectastain ABC kit (rabbit IgG; Vector Laboratories, Burlingame, CA, USA) and the anti-FABP5 (D1A7T) rabbit monoclonal antibody (Cell Signaling Technology, Danvers, MA, USA) diluted with goat blocking serum at 1:200 were used.

Techniques: Expressing

Journal: Aging Cell

Article Title: Accumulation of systematic TPM1 mediates inflammation and neuronal remodeling by phosphorylating PKA and regulating the FABP5/NF‐κB signaling pathway in the retina of aged mice

doi: 10.1111/acel.13566

Figure Lengend Snippet: Systematic TPM1 functions by phosphorylating PKA in aging retinas. (a) ELISA analysis of cAMP in BV2 cells after different treatments. Following exposure to DMSO, rTPM1, FSK, SQ, or FSK + SQ for 24 h, BV2 cells were collected to detect the protein level of cAMP. Data are presented as mean ± SEM and analyzed by one‐way ANOVA with Tukey's multiple comparison test (compared to FSK, * p < 0.05). Five independent experiments were performed. (b–e) Western blot analysis (b) and quantification of TPM1, p‐PKA, and PKA (c–e) in BV2 cells after different treatments. Data are presented as mean ±SEM and analyzed by one‐way ANOVA with Tukey's multiple comparison test (compared with rTPM1, * p < 0.05, ** p < 0.01; compared with FSK, # p < 0.05, ## p < 0.01; SQ vs. FSK + SQ, $ p < 0.05). Five independent experiments were performed. (f–g) Western blot analysis (f) and quantification of TPM1 (g) in BV2 cells after rTPM1 and H89 treatments. Data are presented as mean ± SEM and analyzed by one‐way ANOVA with Tukey's multiple comparison test (compared with rTPM1, ** p < 0.01). (h) ELISA analysis of cAMP in young retinas after YMP or OMP treatment. Data are presented as mean ± SEM, n = 5 mice in each group. (i–n) Western blot analysis (i) and quantification of Adcy2, p‐PKA, PKA, FABP5, and TPM1 (j–n) in young retinas after YMP or OMP treatment. Data are presented as mean ± SEM, n = 4 mice in each group, one‐way ANOVA with Tukey's multiple comparison test. (o) ELISA analysis of cAMP in young retinas after rTPM1 protein or PBS treatment. Data are presented as mean ± SEM, n = 4 mice in each group. (p–u) Western blot analysis (p) and quantification of Adcy2, p‐PKA, PKA, FABP5, and TPM1 proteins (q–u) in young retinas after rTPM1 protein or PBS treatment. Data are presented as mean ± SEM, n = 5 mice in each group, unpaired two‐tailed Student's t test. (v–w) qPCR analysis of TPM1.5, TPM1.9, and TPM1.10 in young (v) or aged (w) retinas after YMP or OMP treatment. Data are presented as mean ± SEM, n = 4 mice in each group, one‐way ANOVA with Tukey's multiple comparison test

Article Snippet: After blocking for 1 h, the membranes were incubated with primary antibodies at the designated concentrations: rabbit anti‐GFAP (Dako, 1:500), rabbit anti‐Iba‐1 (Wako, 1:500) or rat anti‐CD68 (Bio‐rad, 1:500), rabbit anti‐TPM1 (ABclonal, 1:1,000; Invitrogen, 1:1,000), rabbit anti‐ADCY2 (Abcam, 1:1,000; ABclonal, 1:1,000), rabbit anti‐Phospho‐PKA C (Thr197) (Cell Signaling Technology, 1:1,000), rabbit anti‐PKA C‐α (Cell Signaling Technology, 1:1,000), rabbit anti‐ FABP5 (D1A7T) (Cell Signaling Technology, 1:1,000), or mouse anti‐GAPDH (Sigma‐Aldrich,1:1,000; ABclonal, 1:1,000), overnight at 4℃.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Two Tailed Test

Journal: Aging Cell

Article Title: Accumulation of systematic TPM1 mediates inflammation and neuronal remodeling by phosphorylating PKA and regulating the FABP5/NF‐κB signaling pathway in the retina of aged mice

doi: 10.1111/acel.13566

Figure Lengend Snippet: Systematic TPM1 functions through the FABP5/NF‐kB signal pathway. (a) Inflammation‐related DEPs in pairwise comparisons both between young and young‐HP and between aged and aged‐HP. The Y‐axes indicate the Log2 FC of co‐expressed DEPs. (b–e) qPCR analysis of Fabp5, Nfkb2, Rela, and Rel in young retinas after YMP or OMP treatment. Data are presented as mean ± SEM, n = 3 mice in each group, one‐way ANOVA with Tukey's multiple comparison test. (f–i) qPCR analysis of Fabp5, Nfkb2, Rela, and Rel in young retinas after rTPM1 protein or PBS treatment. Data are presented as mean ± SEM, n = 3 mice in each group, unpaired two‐tailed Student's t test. (j–o) Western blot analysis (j) and quantification of Adcy2, p‐PKA, PKA, FABP5, and TPM1 (k–o) proteins in BV2 cells after rTPM1 treatment following the transfection of siFABP5. Data are presented as mean ± SEM and analyzed by one‐way ANOVA with Tukey's multiple comparison test (compared with rTPM1 + siCRT, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.001; compared with rTPM1 + siFABP5, # p < 0.05, ### p < 0.001). (p) ELISA analysis of cAMP in BV2 cells after rTPM1 protein treatment following the transfection of siFABP5. Data are presented as mean ± SEM. (q–t) qPCR analysis of Fabp5, Nfkb2, Rela, and Rel in BV2 cells after rTPM1 treatment following the transfection of siFABP5. Data are presented as mean ± SEM, one‐way ANOVA with Tukey's multiple comparison test. Three independent experiments were performed. (u–v) Western blot analysis (u) and quantification of FABP5 (v) in BV2 cells after rTPM1 and H89 treatment. Data are presented as mean ± SEM. (w) Schematic depicting TPM1‐related signaling pathway. Systematic TPM1 accumulation in young‐HP, OMP‐treated or rTPM1 protein‐treated young retina upregulates endogenous TPM1 by phosphorylating PKA and by activating FABP5/NF‐kB signaling pathway, leading to elevated inflammatory responses and neuronal remodeling in retina aging process

Article Snippet: After blocking for 1 h, the membranes were incubated with primary antibodies at the designated concentrations: rabbit anti‐GFAP (Dako, 1:500), rabbit anti‐Iba‐1 (Wako, 1:500) or rat anti‐CD68 (Bio‐rad, 1:500), rabbit anti‐TPM1 (ABclonal, 1:1,000; Invitrogen, 1:1,000), rabbit anti‐ADCY2 (Abcam, 1:1,000; ABclonal, 1:1,000), rabbit anti‐Phospho‐PKA C (Thr197) (Cell Signaling Technology, 1:1,000), rabbit anti‐PKA C‐α (Cell Signaling Technology, 1:1,000), rabbit anti‐ FABP5 (D1A7T) (Cell Signaling Technology, 1:1,000), or mouse anti‐GAPDH (Sigma‐Aldrich,1:1,000; ABclonal, 1:1,000), overnight at 4℃.

Techniques: Two Tailed Test, Western Blot, Transfection, Enzyme-linked Immunosorbent Assay