stim1 antibody (cdn3h4) - bsa free  (Bio-Techne corporation)

Journal: The Journal of investigative dermatology

Article Title: Plantar Skin Exhibits Altered Physiology, Constitutive Activation of Wound-Associated Phenotypes, and Inherently Delayed Healing.

doi: 10.1016/j.jid.2023.12.016

Figure Lengend Snippet: Figure 5. Differential expression of calcium-binding proteins in human plantar versus nonplantar skin shows disparate presentations of calcium-binding proteins. (a) Human skin samples did not express S100A12 at baseline (not shown) but otherwise showed similar differential expression patterns as porcine skin: specifically, increased staining for S100 proteins and reduced STIM1 in plantar skin. (b) Semiquantitative analysis of STIM1 staining intensity, confirming reduced STIM1 in plantar skin. Bars ¼ 50 mm for dorsal and 25 mm for plantar. Additional information on the presentation of these calcium-binding proteins in human skin is included in Supplementary Figures S6 and S7.

Article Snippet: Primary Antibodies Target Vendor Catalog Number Clone Number Dilution a-SMA Abcam ab 5694 Polyclonal 1:100 Keratin 1 Novus Biologicals NB110-89450 3C10F7 1:800 Keratin 5 Antibodies-online.com ABIN112348 RCK102 1:800 Keratin 6 LSBio LS-C392262-20 SPM269 1:1000 Keratin 9 Abcam ab 171966 EPR10932-62 1:100 Keratin 10 Abcam ab 9026 DE-K10 1:200 Keratin 14 Novus Biologicals NBP3-07600 KRT14/2375 1:400 Keratin 16 Antibodies-online.com ABIN2775315 Polyclonal 1:100 Ki-67 Abcam ab 16667 SP6 1:100 S100A8 MyBioSource MBS2028565 Polyclonal 1:400 S100A12 Novus Biologicals NBP1-86694 Polyclonal 1:250 S100A14 Thermo Fisher Scientific PA5-55666 Polyclonal 1:2500 S100A16 Thermo Fisher Scientific PA5-80987 Polyclonal 1:8000 STIM1 Novus Biologicals NB110-60547 Polyclonal 1:200 Abbreviation: a-SMA, a-smooth muscle actin.

Techniques: Quantitative Proteomics, Binding Assay, Staining

Journal: The Journal of investigative dermatology

Article Title: Plantar Skin Exhibits Altered Physiology, Constitutive Activation of Wound-Associated Phenotypes, and Inherently Delayed Healing.

doi: 10.1016/j.jid.2023.12.016

Figure Lengend Snippet: Figure 6. Functional changes in plantar keratinocytes consistent with diminished STIM1. (a) Store-operated calcium entry in primary porcine trunk and plantar keratinocytes. Trunk keratinocytes (blue curve) showed the typical response: small and transient increase in intracellular Ca2þ upon exposure to TG and then a large spike when extracellular Ca2þ was restored (to 0.5 mM from the previously Ca2þ-free conditions). This response was significantly blunted in plantar keratinocytes (red curve). Data are shown as mean SEM from 7 independent experiments each with cells from a different pig. (b) Peak Ca2þ rise due to Ca2þ

Article Snippet: Primary Antibodies Target Vendor Catalog Number Clone Number Dilution a-SMA Abcam ab 5694 Polyclonal 1:100 Keratin 1 Novus Biologicals NB110-89450 3C10F7 1:800 Keratin 5 Antibodies-online.com ABIN112348 RCK102 1:800 Keratin 6 LSBio LS-C392262-20 SPM269 1:1000 Keratin 9 Abcam ab 171966 EPR10932-62 1:100 Keratin 10 Abcam ab 9026 DE-K10 1:200 Keratin 14 Novus Biologicals NBP3-07600 KRT14/2375 1:400 Keratin 16 Antibodies-online.com ABIN2775315 Polyclonal 1:100 Ki-67 Abcam ab 16667 SP6 1:100 S100A8 MyBioSource MBS2028565 Polyclonal 1:400 S100A12 Novus Biologicals NBP1-86694 Polyclonal 1:250 S100A14 Thermo Fisher Scientific PA5-55666 Polyclonal 1:2500 S100A16 Thermo Fisher Scientific PA5-80987 Polyclonal 1:8000 STIM1 Novus Biologicals NB110-60547 Polyclonal 1:200 Abbreviation: a-SMA, a-smooth muscle actin.

Techniques: Functional Assay

Journal: Journal of Lipid Research

Article Title: Crosstalk between ORMDL3, serine palmitoyltransferase, and 5-lipoxygenase in the sphingolipid and eicosanoid metabolic pathways

doi: 10.1016/j.jlr.2021.100121

Figure Lengend Snippet: Identification of the ORMDL3/5-LO/SPTLC1/SPTLC2 complex. A: ORMDL3-MYC immunoprecipitated from lysates of BMMCL expressing ORMDL3-MYC or empty vectors (control) and activated or not with thapsigargin. The immunoprecipitates were size-fractionated by SDS-PAGE and stained by Coomassie blue. ←5-LO indicates position of 5-LO as identified by mass spectrometry. B: 5-LO interacts with ORMDL3, SPTLC1, and SPTLC2. Twin-Strep-tag (TS) affinity purification from lysates of HEK293FT cells stably transduced with FLAG-ORMDL3 alone or in combination with TS-5-LO and activated for 0, 3, 15, or 25 min with thapsigargin. The proteins were analyzed by immunoblotting using the corresponding antibodies. C: Colocalization of endogenous 5-LO with ORMDLs (left panel) or STIM1 (right panel) in nonactivated (0 min) or activated HMC-1.1 cells for 10 or 30 min with ionomycin. Scale bars represent 5 μm. D: Physical interaction of 5-LO with ORMDL3. FLAP, ORMDL3, LTC4S, and 5-LO were tagged with GST and expressed in bacteria. GST-tagged proteins and GST alone were purified on glutathione-coated beads in the presence of in vitro translate expressing 35 S-5-LO. SDS-PAGE followed by autoradiography was used to detect 35 S-5-LO. The gel was stained with Coomassie brilliant blue (CB staining) to determine the loading levels of GST and GST-tagged proteins. E: SDS-PAGE of lysates from HEK293 cells stably transduced with empty vector (control), murine 5-LO, or murine ORMDL3 developed with the indicated protein-specific antibodies. F and G: LC-ESI-MS/MS analysis of sphingolipids in resting HEK293 cells stably transduced with murine 5-LO (n = 13), murine ORMDL3 (n = 7), or empty vector (control, n = 12). F: Total sphingosines, the sum of C18:1 and C18:0 is calculated. G: The sum of total ceramide fatty acid chain molecular species (including 2-hydroxy ceramide molecular species), derived from d18:1 sphingosine, was calculated. Data in B and D are representative of three independent experiments. Quantitative data in F and G are mean ± SEM, calculated from n, which show numbers of biological replicates of independently transduced cells. P values were determined by one-way ANOVA with Bonferoni post hoc test.

Article Snippet: Antibodies against HA tag (NB600-363) and stromal interaction molecule 1 (STIM1) (NB110-60547S) were obtained from Novus Biologicals.

Techniques: Immunoprecipitation, Expressing, Control, SDS Page, Staining, Mass Spectrometry, Strep-tag, Affinity Purification, Stable Transfection, Transduction, Western Blot, Bacteria, Purification, In Vitro, Autoradiography, Plasmid Preparation, Tandem Mass Spectroscopy, Derivative Assay