Human/Mouse/Rat FABP5/E-FABP Antibody

Detection of Human, Mouse, and Rat FABP5/E‑FABP by Western Blot. Western blot shows lysates of human placenta tissue, mouse liver tissue, rat brain tissue, and rat heart tissue. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/ Rat FABP5/E-FABP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1476) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for FABP5/ E-FABP at approximately 15 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

FABP5/E‑FABP in Mouse Skin Tissue. FABP5/E-FABP was detected in immersion fixed paraffin-embedded sections of mouse skin (ear) tissue using Goat Anti-Human/Mouse/Rat FABP5/E-FABP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1476) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human and Mouse FABP5/E‑FABP by Simple Western^TM. Simple Western lane view shows lysates of human placenta and mouse liver, loaded at 0.2 mg/mL. A specific band was detected for FABP5/E‑FABP at approximately 20 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse/Rat FABP5/E‑FABP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1476). This experiment was conducted under reducing conditions and using the 2-40 kDa separation system.

Western Blot Shows Human FABP5/E‑FABP Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and FABP5/E-FABP knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat FABP5/E-FABP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1476) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for FABP5/E-FABP at approximately 15 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse FABP5/E-FABP by Immunocytochemistry/ Immunofluorescence JunB cKOs display impaired skin homeostasis. a Representative photomicrographs with immunostaining of FABP5 (red), indicative of sebaceous glands, in dorsal back skin from wild type and JunB cKO mice. Nuclei stained with DAPI in blue. Scale bars, 50 µm. b Histology of tail skin from wild type and JunB cKO mice. Scale bars, 50 µm. c Representative photo of colony-forming unit assay from FACS purified HFSCs (CD34+ve alpha 6ItgHi) from wild type and JunB cKO mice back skin that were cultured in vitro for 2 weeks and then stained with Rhodamine B and Nile blue. n = 3 mice per genotype. d FACS analyses from second telogen phase displaying an increased ratio of P2 to P1 hair follicle stem cells (CD34+ve alpha 6ItgHi /CD34+ve alpha 6ItgLow) in unperturbed dorsal skin from 60 days old JunB cKO compared to wild type mice. e Confocal images of whole mount tail epidermis depict differentiated keratinocytes immunostained for the terminal differentiation marker k10 (green) in JunB cKO compared to wild type skin. Nuclei stained with DAPI in blue. Scale bars, 50 µm. E, epidermis; SG, sebaceous gland. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30143626), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse FABP5/E-FABP by Immunohistochemistry JunB expression during skin maturation and upon stress. a Representative microphotographs with double immunostaining for JunB (green) and FABP5 (red), indicative of sebaceous glands, in skin derived from 1, 3, and 5 days old mice. Inset showing magnified view of a sebaceous gland. Scale bars, 50 µm. b Immunostaining of JunB (green) and FABP5 (red) in 60 days old mice. Inset showing magnified view. c Representative microphotographs with double immunostaining of JunB (green) and FABP5 (red) in dorsal skin of 60 days old mice following hair plucking or d after topical TPA application, a potent inducer of proliferation. e Immunostaining of JunB (green) and LRIG1 or CD34 (red) in dorsal skin of 60 days old mice following hair plucking. f Immunostaining of JunB (green) and FABP5 (red) in adult human skin. E, epidermis; D, dermis; HF, hair follicle; SG, sebaceous gland; SD, sebaceous duct; HS, hair shaft. Asterisk indicates hair shaft autofluorescence. Dashed line indicates the epidermal-dermal junction. Scale bars, 50 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30143626), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse FABP5/E-FABP by Immunocytochemistry/ Immunofluorescence De novo formation of sebaceous glands in JunB cKO wounds. a Representative H&E photomicrographs of 30 days old wound restoration tissue depicting ectopic sebaceous glands in JunB cKO mice. Inset showing magnified view. E, epidermis; SG, sebaceous gland. Scale bars, 50 µm. Quantification of de novo sebaceous gland numbers in 30 days old wounds. ***P < 0.001, t-test (n = 3). Graphical sketch summarizes JunB-dependent suppression of epidermal progenitor differentiation towards sebocyte lineage (left bottom panel as opposed to lineage plasticity of epidermal progenitors towards sebocyte differentiation (right bottom panel). b, c Representative microphotographs with immunostaining of SCD1 (red), indicative of sebocyte differentiation, demonstrating different stages of de novo formation of sebaceous glands in 30 days old wounds in JunB cKO mice. Nuclei stained with DAPI in blue. d Representative photomicrographs displaying the formation of elongated sebaceous duct in 90-day-old wound epithelium in JunB cKO mice. Wound tissues were stained either with H & E or the sebaceous glands specific marker FABP5 (red) and the basal layer specific keratin K14 (green). Nuclei stained with DAPI in blue. Scale bars, 50 µm. E, epidermis; SG, sebaceous gland. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30143626), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse FABP5/E-FABP by Immunohistochemistry JunB expression during skin maturation and upon stress. a Representative microphotographs with double immunostaining for JunB (green) and FABP5 (red), indicative of sebaceous glands, in skin derived from 1, 3, and 5 days old mice. Inset showing magnified view of a sebaceous gland. Scale bars, 50 µm. b Immunostaining of JunB (green) and FABP5 (red) in 60 days old mice. Inset showing magnified view. c Representative microphotographs with double immunostaining of JunB (green) and FABP5 (red) in dorsal skin of 60 days old mice following hair plucking or d after topical TPA application, a potent inducer of proliferation. e Immunostaining of JunB (green) and LRIG1 or CD34 (red) in dorsal skin of 60 days old mice following hair plucking. f Immunostaining of JunB (green) and FABP5 (red) in adult human skin. E, epidermis; D, dermis; HF, hair follicle; SG, sebaceous gland; SD, sebaceous duct; HS, hair shaft. Asterisk indicates hair shaft autofluorescence. Dashed line indicates the epidermal-dermal junction. Scale bars, 50 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30143626), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse FABP5/E-FABP by Immunohistochemistry Notch inhibition restored epidermal homeostasis in JunB cKOs. a Cartoon depicting experimental design and administration schedule of the Notch inhibitor DBZ or vehicle in JunB cKO mice. b Representative H&E staining and c immunostaining for the sebaceous gland specific marker FABP5 demonstrates the absence of de novo sebaceous gland formation in newly formed wound epithelium of the Notch inhibitor DBZ treated JunB cKO mice as opposed to vehicle treated group (n = 3). Scale bars, 50 µm. d Representative microphotographs with immunostaining of differentiated keratinocytes marker K10 (red) and undifferentiated keratinocytes marker K14 (green), in DBZ treated JunB cKO mice compared to vehicle treated group (n = 3). Nuclei stained with DAPI in blue. Scale bars, 50 µm. e Representative microphotographs with immunostaining of proliferation marker Ki67 (green) and FABP5 (red) and f quantification, in DBZ treated JunB cKO mice as opposed to vehicle treated group. ***P < 0.001, t-test (n = 3). Nuclei stained with DAPI in blue. Scale bars, 50 µm. g TEWL, which is inversely related with epidermal barrier function, was measured on the dorsal skin of DBZ treated JunB cKO mice and vehicle treated group (n = 3). ***P < 0.001, t-test. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30143626), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse FABP5/E-FABP by Immunohistochemistry Notch inhibition restored epidermal homeostasis in JunB cKOs. a Cartoon depicting experimental design and administration schedule of the Notch inhibitor DBZ or vehicle in JunB cKO mice. b Representative H&E staining and c immunostaining for the sebaceous gland specific marker FABP5 demonstrates the absence of de novo sebaceous gland formation in newly formed wound epithelium of the Notch inhibitor DBZ treated JunB cKO mice as opposed to vehicle treated group (n = 3). Scale bars, 50 µm. d Representative microphotographs with immunostaining of differentiated keratinocytes marker K10 (red) and undifferentiated keratinocytes marker K14 (green), in DBZ treated JunB cKO mice compared to vehicle treated group (n = 3). Nuclei stained with DAPI in blue. Scale bars, 50 µm. e Representative microphotographs with immunostaining of proliferation marker Ki67 (green) and FABP5 (red) and f quantification, in DBZ treated JunB cKO mice as opposed to vehicle treated group. ***P < 0.001, t-test (n = 3). Nuclei stained with DAPI in blue. Scale bars, 50 µm. g TEWL, which is inversely related with epidermal barrier function, was measured on the dorsal skin of DBZ treated JunB cKO mice and vehicle treated group (n = 3). ***P < 0.001, t-test. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30143626), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse FABP5/E-FABP by Immunohistochemistry JunB expression during skin maturation and upon stress. a Representative microphotographs with double immunostaining for JunB (green) and FABP5 (red), indicative of sebaceous glands, in skin derived from 1, 3, and 5 days old mice. Inset showing magnified view of a sebaceous gland. Scale bars, 50 µm. b Immunostaining of JunB (green) and FABP5 (red) in 60 days old mice. Inset showing magnified view. c Representative microphotographs with double immunostaining of JunB (green) and FABP5 (red) in dorsal skin of 60 days old mice following hair plucking or d after topical TPA application, a potent inducer of proliferation. e Immunostaining of JunB (green) and LRIG1 or CD34 (red) in dorsal skin of 60 days old mice following hair plucking. f Immunostaining of JunB (green) and FABP5 (red) in adult human skin. E, epidermis; D, dermis; HF, hair follicle; SG, sebaceous gland; SD, sebaceous duct; HS, hair shaft. Asterisk indicates hair shaft autofluorescence. Dashed line indicates the epidermal-dermal junction. Scale bars, 50 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30143626), licensed under a CC-BY license. Not internally tested by R&D Systems.