Review

pcad antibody (Novus Biologicals)

Novus Biologicals is a verified supplier

Novus Biologicals manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Novus Biologicals pcad antibody
Pcad Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pcad antibody/product/Novus Biologicals
Average 90 stars, based on 1 article reviews

pcad antibody - by Bioz Stars, 2026-04

90/100 stars

Images

Image Search Results

Journal: Nature protocols

Article Title: Generation and characterization of hair-bearing skin organoids from human pluripotent stem cells

doi: 10.1038/s41596-022-00681-y

Figure Lengend Snippet: a-e, See Table 1 for further descriptions. Representative IHC images showing key checkpoints throughout the differentiation procedure of WA25 hESC-derived skin organoids. a, Day 18 image representing the ECAD+TFAP2A+ epithelium and TFAP2A+ CNC or mesenchymal cells surrounding the aggregate; the presence of these cell populations should be checked between days 6–20 of differentiation. b, Day 55 images showing TFAP2A+KRT5+ and KRT17+KRT5+ basal layer, TFAP2A+ intermediate layer, and KRT17+ periderm layer; the periderm layer is detectable only at early stages of differentiation, prior to formation of the granular and cornified layers; the periderm layer is visible in organoids around days 40–75 of differentiation. c, Overall-view image of day 70 skin cysts; the images show the major layers of skin that are required to form the skin, the epidermal and the dermal layers, and the initiating hair germs; the basal layer of skin is highlighted by KRT5+KRT15+ and CD49f+ fluorescence signals; the periderm layer is visualized by KRT15; the dermal layer (fibroblasts) is visualized by PDGFRα, and NC cell-derived mesenchymal cells within the population express P75; the SOX2+ cells represent dermal condensates at the tip of the hair germs; the initial hair placode and germ formations can be observed starting around day 55 of differentiation. PD, periderm; DC, dermal condensate. d,e, Day 75 high-magnification images representing PCAD+ hair placodes, PCAD+EDAR+LHX2+ hair germs, and PCAD+LHX2+ hair pegs; SOX2+ cells represent dermal condensates of hair germs and dermal papillae of hair pegs. f, A representative IHC image of a day 140 skin organoid. The endogenous green fluorescence from the DSP-GFP cell line visualizes epithelium of the skin cyst in the center and the hair follicles protruding from the surface of the cyst. TUJ1+ neurons are wrapping around and innervating the epithelium and the hair follicles. The skin organoids reach the lanugo-like mature stage around day 120 of differentiation. g, Representative darkfield images of a day 125 WA25 hESC-derived skin organoid (left) and dermal view of 18-week human fetal facial skin tissue (right). Skin organoids at days 120–140 resemble the mid-second trimester fetal skin tissue with (pigmented) hair follicles and adipocytes. h,i, Representative whole-mount immunostaining images of hair follicles with dermal papillae and melanocytes in a day 85 (h, left and middle) and a day 120 (i) WA25 hESC-derived skin organoids. KRT5 visualizes epithelium, outer root sheath (ORS) of hair follicles and newly forming hair germs (h, middle). SOX2 marks for melanocytes or Merkel cells present in the ORS of hair follicles and on the epithelium (h, right). MITF also specifies melanocytes in the ORS and on the epithelium. Dermal papillae of the hair follicles are also visualized by P75 (i). j, A representative whole-mount immunostaining image of a day 110 skin organoid hair follicles. The hair follicles contain a bulge region where KRT20+ touch-sensing Merkel cells are present. NEFH+ sensory neurons innervate the upper bulge region near Merkel cells. k, Representative brightfield images of plucked hairs from human fetal facial tissue at 18 weeks of gestation, adult male’s cheek (beard), skin organoid xenograft, and DSP skin organoid at day 190 of differentiation. Insets present a magnified area indicated with dash boxes. The medulla is only present in the adult beard. The medulla layer is not visible in xenograft hairs, either pigmented or non-pigmented. Darker hairs from a xenograft and a DSP skin organoid appear to contain pigmented cells that are scattered throughout the cortex, but no sign of medulla is detectable in the center of the hair shaft. See ref. 25 for additional images. The images are taken at the magnifications as follows: 200X (20X microscope objective × 10X eyepiece; b, d, e, h’); 100X (10X microscope objective × 10X eyepiece; a, c, h (left), i, and j); 50X (5X microscope objective × 10X eyepiece; g); 40X (4X microscope objective × 10X eyepiece; f and k (xenografts and organoid hairs)); 20X (2X microscope objective × 10X eyepiece; k (fetal hair and adult beard)). Scale bars, 500 μm (f, g, and k); 200 μm (a); 100 μm (b-e, and j); 50 μm (h (left) and i); 30 μm (h’).

Article Snippet: P-cadherin (PCAD) , Thermo Fisher Scientific , 32-4000 , AB_2533077 https://scicrunch.org/resolver/RRID:AB_2533077 , Mouse , IgG 1 , 1:50 , Hair placodes, germs, and pegs.

Techniques: Derivative Assay, Fluorescence, Immunostaining, Microscopy

Journal: Nature protocols

Article Title: Generation and characterization of hair-bearing skin organoids from human pluripotent stem cells

doi: 10.1038/s41596-022-00681-y

Figure Lengend Snippet: Major checkpoints

Article Snippet: P-cadherin (PCAD) , Thermo Fisher Scientific , 32-4000 , AB_2533077 https://scicrunch.org/resolver/RRID:AB_2533077 , Mouse , IgG 1 , 1:50 , Hair placodes, germs, and pegs.

Techniques: Imaging

Journal: Nature protocols

Article Title: Generation and characterization of hair-bearing skin organoids from human pluripotent stem cells

doi: 10.1038/s41596-022-00681-y

Figure Lengend Snippet: Primary antibodies

Article Snippet: P-cadherin (PCAD) , Thermo Fisher Scientific , 32-4000 , AB_2533077 https://scicrunch.org/resolver/RRID:AB_2533077 , Mouse , IgG 1 , 1:50 , Hair placodes, germs, and pegs.

Techniques: Isotope Dilution, Marker

(A) The de novo pyrimidine synthesis pathway. The rate-limiting trifunctional enzyme CAD is phosphorylated and activated by S6K1, which is in turn phosphorylated and activated by mTORC1. The CAD protein catalyzes the first three steps of this pathway and is inhibited by the small molecule PALA (shown in red). (B) Western blot analysis of CD8+ T cells isolated from spleens of naïve WT mice and stimulated with anti-CD3/28 antibodies for indicated time periods with or without rapamycin (100 nM). (C) Western blot analysis of CD8+ T cells isolated from spleens of naïve WT, Rptor−/− and Rictor−/− mice stimulated with anti-CD3/28 antibodies for indicated three hours. (D) Flow cytometric analysis of WT, Rptor−/− and Rictor−/− mice for pCAD. CD8+ T cells from WT mice were analyzed as either unstimulated (grey) or stimulated using anti-CD3/28 antibodies for three hours with 100 nM rapamycin (black) or without rapamycin (red). Rptor−/− (blue) and Rictor−/− (green) CD8+ T cells were also stimulated for three hours. (E) Western blot analysis of CD8+ T cells isolated from spleens of naïve WT mice and stimulated with anti-CD3/28 antibodies for 36 and 72 hours. (F) Western blot analysis of CD8+ T cells isolated from spleens of naïve C57BL/6J mice and stimulated with anti-CD3/28 antibodies for 48 hours, then expanded in media containing IL-2 or IL-7 + IL-15 for an additional five days. Data in (B) includes representative image and densitometry of fold increase in signal relative to actin across three experiments. Data in (C) includes representative image and densitometry of signal relative to actin across three experiments. Data in (D) included representative image and depicts fold change in MFI over naïve CD8+cells across three experiments. Data in (E) and (F) include representative image and densitometry of signal ratio of phosphoprotein to total protein relative to actin across three experiments. Data in (B), (E), and (F) analyzed by two-tailed t test. Data in (C) and (D) analyzed by one-way ANOVA followed by Tukey’s HSD test. *P <0.05, ns = not significant.

Journal: Science immunology

Article Title: Persistent CAD activity in memory CD8 + T cells supports rRNA synthesis and ribosomal biogenesis required at rechallenge

doi: 10.1126/sciimmunol.abh4271

Figure Lengend Snippet: (A) The de novo pyrimidine synthesis pathway. The rate-limiting trifunctional enzyme CAD is phosphorylated and activated by S6K1, which is in turn phosphorylated and activated by mTORC1. The CAD protein catalyzes the first three steps of this pathway and is inhibited by the small molecule PALA (shown in red). (B) Western blot analysis of CD8+ T cells isolated from spleens of naïve WT mice and stimulated with anti-CD3/28 antibodies for indicated time periods with or without rapamycin (100 nM). (C) Western blot analysis of CD8+ T cells isolated from spleens of naïve WT, Rptor−/− and Rictor−/− mice stimulated with anti-CD3/28 antibodies for indicated three hours. (D) Flow cytometric analysis of WT, Rptor−/− and Rictor−/− mice for pCAD. CD8+ T cells from WT mice were analyzed as either unstimulated (grey) or stimulated using anti-CD3/28 antibodies for three hours with 100 nM rapamycin (black) or without rapamycin (red). Rptor−/− (blue) and Rictor−/− (green) CD8+ T cells were also stimulated for three hours. (E) Western blot analysis of CD8+ T cells isolated from spleens of naïve WT mice and stimulated with anti-CD3/28 antibodies for 36 and 72 hours. (F) Western blot analysis of CD8+ T cells isolated from spleens of naïve C57BL/6J mice and stimulated with anti-CD3/28 antibodies for 48 hours, then expanded in media containing IL-2 or IL-7 + IL-15 for an additional five days. Data in (B) includes representative image and densitometry of fold increase in signal relative to actin across three experiments. Data in (C) includes representative image and densitometry of signal relative to actin across three experiments. Data in (D) included representative image and depicts fold change in MFI over naïve CD8+cells across three experiments. Data in (E) and (F) include representative image and densitometry of signal ratio of phosphoprotein to total protein relative to actin across three experiments. Data in (B), (E), and (F) analyzed by two-tailed t test. Data in (C) and (D) analyzed by one-way ANOVA followed by Tukey’s HSD test. *P <0.05, ns = not significant.

Article Snippet: Primary antibodies included CD8α (53–6.7), CD44 (IM7), IL-2 (JES6-5H4), TNF-α (MP6-XT22), IFN-γ (XMG1.2), CD45RO (UCHL1), CD45RA (HI100), CD3 (UCHT1), IFN-γ (B27), and TNF-α ((RUO(GMP)) from BD Biosciences, CD62L (MEL-14) from BioLegend, anti-CAD (11933S), anti pCAD (S1859, 12662S), anti-S6 (2217), anti-pS6 (S240/244, D68F8), anti S6K (9202), anti-pS6K (T389, 9205S), anti-RPL5 (14568S), and anti-β-Actin (D6A8, 8457) from Cell Signaling, Fixable Viability Dye eFluor 780 (65-0865-14) from eBioscience, anti-HA Alexa Fluor 488 conjugated (F-7, sc-7392) from Santa Cruz Biotechnology, and anti-RPL22 (PA5-97192) from ThermoFisher.

Techniques: Western Blot, Isolation, Two Tailed Test

(A) Flow cytometric analysis of pCAD signal in naïve cells (CD8+CD44−CD62L+) and CM cells (CD8+CD44+CD62L+) isolated from the spleens of uninfected WT mice. (B) Flow cytometric analysis of pCAD in naïve and antigen-specific CM CD8+ cells. CD8+ cells were isolated from spleens of naïve OT-I mice and adoptively transferred into WT hosts which were immediately infected with Vac-OVA with splenocytes harvested forty five days later. (C) Flow cytometric analysis of pCAD and pS6 during viral infection. Experiment was performed as in (B) with splenocytes harvested eight days and fifteen days post-infection. (D) Schematic of in vivo rapamycin administration. Mice were administered Vehicle, Low Dose (75 μg/kg) or High Dose (3 mg/kg) rapamycin for Day −1 to Day 8 of Listeria-OVA infection with adoptive transfer of cells freshly isolated from naïve OT-I mice on Day 0. Splenocytes were harvested on Day 35 of infection for analysis by flow cytometry. (E) Summary pCAD intensity and Thy1.1+ data from splenocytes in experiments described in (D) across three replicates. Data in (A) and (B) depict representative gating and fluorescent staining alongside summary MFI values ± SD across three independent experiments normalized to naïve CD8+ cell values within each experiment analyzed by paired two-tailed t-test (n=5 mice per experiment). Data in (C) show mean fluorescence intensity values across three independent experiments normalized to naïve values within each experiment analyzed by paired two-tailed t-test (n=3 mice per experiment). Data in (E) depict representative gating and percentage of Thy1.1+ cells among CD8+ cells and pCAD MFI values ± SD across three independent experiments normalized to vehicle treated CD8+ cell values within each experiment analyzed by one-way ANOVA followed by Tukey’s HSD test. *P <0.05, **P<0.01, ***P<0.001, ****P<0.0001, ns = not significant.

Journal: Science immunology

Article Title: Persistent CAD activity in memory CD8 + T cells supports rRNA synthesis and ribosomal biogenesis required at rechallenge

doi: 10.1126/sciimmunol.abh4271

Figure Lengend Snippet: (A) Flow cytometric analysis of pCAD signal in naïve cells (CD8+CD44−CD62L+) and CM cells (CD8+CD44+CD62L+) isolated from the spleens of uninfected WT mice. (B) Flow cytometric analysis of pCAD in naïve and antigen-specific CM CD8+ cells. CD8+ cells were isolated from spleens of naïve OT-I mice and adoptively transferred into WT hosts which were immediately infected with Vac-OVA with splenocytes harvested forty five days later. (C) Flow cytometric analysis of pCAD and pS6 during viral infection. Experiment was performed as in (B) with splenocytes harvested eight days and fifteen days post-infection. (D) Schematic of in vivo rapamycin administration. Mice were administered Vehicle, Low Dose (75 μg/kg) or High Dose (3 mg/kg) rapamycin for Day −1 to Day 8 of Listeria-OVA infection with adoptive transfer of cells freshly isolated from naïve OT-I mice on Day 0. Splenocytes were harvested on Day 35 of infection for analysis by flow cytometry. (E) Summary pCAD intensity and Thy1.1+ data from splenocytes in experiments described in (D) across three replicates. Data in (A) and (B) depict representative gating and fluorescent staining alongside summary MFI values ± SD across three independent experiments normalized to naïve CD8+ cell values within each experiment analyzed by paired two-tailed t-test (n=5 mice per experiment). Data in (C) show mean fluorescence intensity values across three independent experiments normalized to naïve values within each experiment analyzed by paired two-tailed t-test (n=3 mice per experiment). Data in (E) depict representative gating and percentage of Thy1.1+ cells among CD8+ cells and pCAD MFI values ± SD across three independent experiments normalized to vehicle treated CD8+ cell values within each experiment analyzed by one-way ANOVA followed by Tukey’s HSD test. *P <0.05, **P<0.01, ***P<0.001, ****P<0.0001, ns = not significant.

Techniques: Isolation, Infection, In Vivo, Adoptive Transfer Assay, Flow Cytometry, Staining, Two Tailed Test, Fluorescence

(A-C) CD8+ cells isolated from WT mice were stimulated with anti-CD3/28 antibodies for 24 hours before introduction of indicated retroviral constructs with cells analyzed on Day 6 of culture. (A) Western blot analysis of pCAD (S1859) and total CAD levels in CD8+ cells following retroviral transduction with EV-MigR1 or CAD-MigR1 constructs. (B) Percentage M+1 peak data for de novo pyrimidine synthesis metabolites measured by LC/MS between EV-MigR1 and CAD-MigR1 cells. On Day 6 of culture, freshly-isolated naïve, EV-MigR1 and CAD-MigR1-transduced cells were rested in culture media before transfer to glutamine-free media supplemented with 4 mM amide-labeled 15-N glutamine, with a portion of each condition stimulated with anti-CD3/28 antibodies. (C) Summary data of cytokine response of retrovirally-transduced cells across three experiments generated as in (B) following 4 hours stimulation with PMA/ionomycin. Data gated on live CD8+GFP+ cells with percentage IFNγ+TNFα+ cells reported. (D) Schematic of adoptive transfer experiments with previously activated P14 retrovirally-transduced cells. (E) Percentage GFP+ before transfer and at Day 4 and Day 6 following adoptive transfer and infection with LCMV-Armstrong. Data in (A) depict representative western blot images and densitometry quantification of indicated protein relative to actin from three independent experiments analyzed by two-tailed t test. Data in (B) are mean ± SD of triplicate samples representative of two independent experiments analyzed by two-way ANOVA followed by Tukey’s HSD test. Data in (C) depicts percentage double positive cells across three independent experiments analyzed by two-tailed t test. Data in (E) depict representative gating strategies and GFP+ ± SD analyzed with two-tailed t test at each indicated time point (n=5 mice per condition). *P <0.05, **P<0.01, ***P<0.001.

Journal: Science immunology

Article Title: Persistent CAD activity in memory CD8 + T cells supports rRNA synthesis and ribosomal biogenesis required at rechallenge

doi: 10.1126/sciimmunol.abh4271

Figure Lengend Snippet: (A-C) CD8+ cells isolated from WT mice were stimulated with anti-CD3/28 antibodies for 24 hours before introduction of indicated retroviral constructs with cells analyzed on Day 6 of culture. (A) Western blot analysis of pCAD (S1859) and total CAD levels in CD8+ cells following retroviral transduction with EV-MigR1 or CAD-MigR1 constructs. (B) Percentage M+1 peak data for de novo pyrimidine synthesis metabolites measured by LC/MS between EV-MigR1 and CAD-MigR1 cells. On Day 6 of culture, freshly-isolated naïve, EV-MigR1 and CAD-MigR1-transduced cells were rested in culture media before transfer to glutamine-free media supplemented with 4 mM amide-labeled 15-N glutamine, with a portion of each condition stimulated with anti-CD3/28 antibodies. (C) Summary data of cytokine response of retrovirally-transduced cells across three experiments generated as in (B) following 4 hours stimulation with PMA/ionomycin. Data gated on live CD8+GFP+ cells with percentage IFNγ+TNFα+ cells reported. (D) Schematic of adoptive transfer experiments with previously activated P14 retrovirally-transduced cells. (E) Percentage GFP+ before transfer and at Day 4 and Day 6 following adoptive transfer and infection with LCMV-Armstrong. Data in (A) depict representative western blot images and densitometry quantification of indicated protein relative to actin from three independent experiments analyzed by two-tailed t test. Data in (B) are mean ± SD of triplicate samples representative of two independent experiments analyzed by two-way ANOVA followed by Tukey’s HSD test. Data in (C) depicts percentage double positive cells across three independent experiments analyzed by two-tailed t test. Data in (E) depict representative gating strategies and GFP+ ± SD analyzed with two-tailed t test at each indicated time point (n=5 mice per condition). *P <0.05, **P<0.01, ***P<0.001.

Techniques: Isolation, Retroviral, Construct, Western Blot, Transduction, Liquid Chromatography with Mass Spectroscopy, Labeling, Generated, Adoptive Transfer Assay, Infection, Two Tailed Test

(A) Flow cytometric analysis of pCAD in naïve (CD45RA+CD45RO−) and central memory (CD45RA−CD45RO+CCR7+) human CD3+CD8+ cells. PBMCs were freshly isolated from healthy donor blood before fixation and analysis. (B) Flow cytometric analysis of pCAD levels in Cad-overexpressing (pcDNA3-CAD) or empty vector-expressing human (pcDNA3-EV) cells. Data shown are gated on live CD3+CD8+HA+ cells. (C) Schematic for overexpression of HA-tagged CAD construct in cultured human CD8+ T cells. (D) Flow cytometric analysis of IFNγ and TNFα production by CD8+ cells with and without Cad overexpression following indicated drug treatment. Cells were exposed to drug for 24 hours before drug removal and four hours of restimulation with PMA/ionomycin. Data shown are gated on CD3+CD8+HA+ cells. (E) Western blot of ribosomal proteins in naïve and memory CD8+ T cells. Human PBMCs were thawed and magnetically sorted into naive (CD8+CD45RO−) and memory (CD8+CD45RO+) populations for lysis and analysis. Data in (A) representative of three healthy donors and is depicted alongside representative gating. Data in (B), (D), and (E) depict representative images as well as summary data across three independent experiments with three frozen PBMC aliquots from three separate healthy donors with data in (B) and (E) analyzed by two-tailed t test and data in (D) analyzed by two-way ANOVA followed by Tukey’s HSD test. *P <0.05, **P<0.01, ***P<0.001.

Journal: Science immunology

Article Title: Persistent CAD activity in memory CD8 + T cells supports rRNA synthesis and ribosomal biogenesis required at rechallenge

doi: 10.1126/sciimmunol.abh4271

Figure Lengend Snippet: (A) Flow cytometric analysis of pCAD in naïve (CD45RA+CD45RO−) and central memory (CD45RA−CD45RO+CCR7+) human CD3+CD8+ cells. PBMCs were freshly isolated from healthy donor blood before fixation and analysis. (B) Flow cytometric analysis of pCAD levels in Cad-overexpressing (pcDNA3-CAD) or empty vector-expressing human (pcDNA3-EV) cells. Data shown are gated on live CD3+CD8+HA+ cells. (C) Schematic for overexpression of HA-tagged CAD construct in cultured human CD8+ T cells. (D) Flow cytometric analysis of IFNγ and TNFα production by CD8+ cells with and without Cad overexpression following indicated drug treatment. Cells were exposed to drug for 24 hours before drug removal and four hours of restimulation with PMA/ionomycin. Data shown are gated on CD3+CD8+HA+ cells. (E) Western blot of ribosomal proteins in naïve and memory CD8+ T cells. Human PBMCs were thawed and magnetically sorted into naive (CD8+CD45RO−) and memory (CD8+CD45RO+) populations for lysis and analysis. Data in (A) representative of three healthy donors and is depicted alongside representative gating. Data in (B), (D), and (E) depict representative images as well as summary data across three independent experiments with three frozen PBMC aliquots from three separate healthy donors with data in (B) and (E) analyzed by two-tailed t test and data in (D) analyzed by two-way ANOVA followed by Tukey’s HSD test. *P <0.05, **P<0.01, ***P<0.001.

Techniques: Isolation, Plasmid Preparation, Expressing, Over Expression, Construct, Cell Culture, Western Blot, Lysis, Two Tailed Test

Review

Structured Review

Images

Similar Products

Image Search Results