rabbit anti human ca1 antibody  (Novus Biologicals)

Journal: Cellular and molecular gastroenterology and hepatology

Article Title: Decreased Colonic Activin Receptor-Like Kinase 1 Disrupts Epithelial Barrier Integrity in Patients With Crohn's Disease.

doi: 10.1016/j.jcmgh.2020.06.005

Figure Lengend Snippet: Figure 5. BMP9–ALK1 signaling is associated with epithelial cell differentiation toward colonocytes. (A) Lineage-specific gene expression in NIBD patient-derived colonic epithelial cell monolayers. Expanded cells were cultured in expansion media in the presence or absence of BMP9 and ALK1–Fc chimera protein. Each gene expression was normalized to RPLP0 (N ¼ 6 per group). (B) CA1 protein expression in NIBD patient-derived colonic epithelial monolayers (N ¼ 4 per group). (C) Colonocyte marker expression in colonic epithelial cells isolated from CD patients (N ¼ 15) and NIBD controls (N ¼ 12). Each gene expression was normalized to GAPDH. (D) CA1 expression by immunohistochemistry in the colonic mucosa of NIBD controls (left) and CD patients (right). (E) CA1 protein expression in the colonic mucosa of NIBD controls and CD patients (N ¼ 5 per group). *P < .05, **P < .01, ***P < .001, and ****P < .0001. P values were determined by the (A and B) Friedman test followed by the Dunn multiple comparison test, or the (C and E) Mann–Whitney test.

Article Snippet: Rabbit anti-human CA1 antibody was obtained from Novus Biologicals (NBP1-88191).

Techniques: Cell Differentiation, Gene Expression, Derivative Assay, Cell Culture, Expressing, Marker, Isolation, Immunohistochemistry, Comparison, MANN-WHITNEY

Journal: eLife

Article Title: Increased expression of heme-binding protein 1 early in Alzheimer's disease is linked to neurotoxicity

doi: 10.7554/eLife.47498

Figure Lengend Snippet: ( A, B, C ) Activation of biological processes at different stages of the disease assessed by Ingenuity Pathway Analysis (IPA). Heat maps represent activation z-score change over the course of disease progression and indicate pathways that are activated at 6M ( A ), 12M ( B ) and 18M ( C ). Data were obtained from four biological replicates per group for each time point. Z-score is calculated based on experimental protein expression data (log 2 AD/control ratio) and the theoretical information stored in the IPA Knowledge Base. Positive value of z-score indicates an activation of biological pathway or function. Distribution of the quantified proteins at 2M ( D ), 6M ( E ), 12M ( F ) and 18M ( G ) based on log 2 ratio AD/Control and p-value (t-test) by time point. The pie charts represent the number of quantified non-regulated proteins (grey), significantly different proteins between 3×Tg-AD and control samples, t-test p-value 0.05 (pink) and significantly regulated proteins with more than 50% expression change in comparison to the control (red). ( H–I ) Dynamics of protein expression over the course of AD progression for a selection of the most regulated proteins based on their function. Proteins involved in mRNA processing and transport ( H ) that are upregulated over time and serine protease inhibitors ( I ) that are downregulated. ( J–M ) Putative presymptomatic protein markers of the disease. ( J ) Top 10 significantly up- and downregulated proteins in 3×Tg-AD mice at presymptomatic time point (2M). ( K ) Immunoblot analysis of most regulated hits. Soluble fractions of brain proteins were analyzed from four 2-month-old control and 3×Tg-AD mice animals, respectively. Hebp1 and Glo1 levels were consistently elevated in the transgenic animals as compared to wild type controls. Ca1 levels were reduced in transgenic animals. Presymptomatic markers that remain up- ( L ) or downregulated ( M ) across the AD progression. 10.7554/eLife.47498.007 Figure 2—source data 1. Full list of quantified proteins in the soluble brain fraction of 3×Tg-AD and wild-type mice.

Article Snippet: The following primary antibodies were used for immunoblotting: rabbit Hebp1 (1:1000, Invitrogen, PA5-30609), mouse Glo1 (1:1000, GeneTex, Irvine, CA, GTX628890), rabbit CA1 (1:250, Novus Biologicals, Abingdon, UK, NBP1-88191), mouse α-tubulin (1:5000, Synaptic Systems, Göttingen, Germany, 302 211), rabbit β-actin (1:5000, Synaptic Systems, 251 003), rabbit GFP (1:5000, Synaptic Systems, 132 002), mouse Rab5 (1:1000, Synaptic Systems, 108 111), rabbit Rab6 (1:1000, Synaptic Systems, 273 003), rabbit Lamp1 (1:500, Abcam, ab24170), mouse Mic60 (1:1000, Abcam, ab110329), rabbit Cox4 (1:1000, Synaptic Systems, 298 002), rabbit Cyt C (1:1000, Cell Signaling, Beverly, MA, USA, 11940S), rabbit caspase 9 (1:1000, Abcam, Cambridge, UK, ab185719), mouse Sodium Potassium ATPase, subunit α1 (1:1000, Abcam, Cambridge, UK, ab7671), mouse syntaxin 1 (1:1000, Synaptic Systems, 110 001), mouse VAMP2 (1:10000, Synaptic Systems, 104 211), rabbit phospho-tau (Ser400;Thr403;Ser404) (1:1000, Cell Signaling, Beverly, MA, USA, 11837S).

Techniques: Activation Assay, Biomarker Discovery, Expressing, Control, Comparison, Selection, Western Blot, Transgenic Assay

Journal: eLife

Article Title: Increased expression of heme-binding protein 1 early in Alzheimer's disease is linked to neurotoxicity

doi: 10.7554/eLife.47498

Figure Lengend Snippet: Identified presymptomatic brain markers of AD in this study.

Article Snippet: The following primary antibodies were used for immunoblotting: rabbit Hebp1 (1:1000, Invitrogen, PA5-30609), mouse Glo1 (1:1000, GeneTex, Irvine, CA, GTX628890), rabbit CA1 (1:250, Novus Biologicals, Abingdon, UK, NBP1-88191), mouse α-tubulin (1:5000, Synaptic Systems, Göttingen, Germany, 302 211), rabbit β-actin (1:5000, Synaptic Systems, 251 003), rabbit GFP (1:5000, Synaptic Systems, 132 002), mouse Rab5 (1:1000, Synaptic Systems, 108 111), rabbit Rab6 (1:1000, Synaptic Systems, 273 003), rabbit Lamp1 (1:500, Abcam, ab24170), mouse Mic60 (1:1000, Abcam, ab110329), rabbit Cox4 (1:1000, Synaptic Systems, 298 002), rabbit Cyt C (1:1000, Cell Signaling, Beverly, MA, USA, 11940S), rabbit caspase 9 (1:1000, Abcam, Cambridge, UK, ab185719), mouse Sodium Potassium ATPase, subunit α1 (1:1000, Abcam, Cambridge, UK, ab7671), mouse syntaxin 1 (1:1000, Synaptic Systems, 110 001), mouse VAMP2 (1:10000, Synaptic Systems, 104 211), rabbit phospho-tau (Ser400;Thr403;Ser404) (1:1000, Cell Signaling, Beverly, MA, USA, 11837S).

Techniques: Binding Assay, Clinical Proteomics, Membrane

Journal: eLife

Article Title: Increased expression of heme-binding protein 1 early in Alzheimer's disease is linked to neurotoxicity

doi: 10.7554/eLife.47498

Figure Lengend Snippet: ( A ) Expression of Hebp1 in 12-month-old control and 3×Tg-AD mice by brain region. ( B ) Hebp1 immunostaining of the fronto-temporal cortex depicting primary motor and somatosensory areas and hippocampus (coronal sections). CA1 region is marked with the white dashed line. ( C ) Co-staining of Hebp1 with markers of CA1 and dentate gyrus neurons (Ctip2), astrocytes (GFAP) and microglia (IBA-1) in the hippocampus of 3×Tg-AD mice. Hepb1 is expressed predominantly in Ctip2-positive cells of hippocampus (neurons). All images were acquired from 12-month-old control or 3×Tg-AD mice. Scale bar is 100 µm. All data shown are representative of results obtained from three independent experiments.

Article Snippet: The following primary antibodies were used for immunoblotting: rabbit Hebp1 (1:1000, Invitrogen, PA5-30609), mouse Glo1 (1:1000, GeneTex, Irvine, CA, GTX628890), rabbit CA1 (1:250, Novus Biologicals, Abingdon, UK, NBP1-88191), mouse α-tubulin (1:5000, Synaptic Systems, Göttingen, Germany, 302 211), rabbit β-actin (1:5000, Synaptic Systems, 251 003), rabbit GFP (1:5000, Synaptic Systems, 132 002), mouse Rab5 (1:1000, Synaptic Systems, 108 111), rabbit Rab6 (1:1000, Synaptic Systems, 273 003), rabbit Lamp1 (1:500, Abcam, ab24170), mouse Mic60 (1:1000, Abcam, ab110329), rabbit Cox4 (1:1000, Synaptic Systems, 298 002), rabbit Cyt C (1:1000, Cell Signaling, Beverly, MA, USA, 11940S), rabbit caspase 9 (1:1000, Abcam, Cambridge, UK, ab185719), mouse Sodium Potassium ATPase, subunit α1 (1:1000, Abcam, Cambridge, UK, ab7671), mouse syntaxin 1 (1:1000, Synaptic Systems, 110 001), mouse VAMP2 (1:10000, Synaptic Systems, 104 211), rabbit phospho-tau (Ser400;Thr403;Ser404) (1:1000, Cell Signaling, Beverly, MA, USA, 11837S).

Techniques: Expressing, Control, Immunostaining, Staining

Journal: eLife

Article Title: Increased expression of heme-binding protein 1 early in Alzheimer's disease is linked to neurotoxicity

doi: 10.7554/eLife.47498

Figure Lengend Snippet: Immunostaining of Hebp1 in the hippocampal region of 3×Tg-AD (27-month-old) and control (24-month-old) mice. Boxed regions in CA1 and dentate gyrus show representative regions where Hebp1 expression was elevated in 3×Tg-AD but not control mice. Scale bar is 500 µm.

Article Snippet: The following primary antibodies were used for immunoblotting: rabbit Hebp1 (1:1000, Invitrogen, PA5-30609), mouse Glo1 (1:1000, GeneTex, Irvine, CA, GTX628890), rabbit CA1 (1:250, Novus Biologicals, Abingdon, UK, NBP1-88191), mouse α-tubulin (1:5000, Synaptic Systems, Göttingen, Germany, 302 211), rabbit β-actin (1:5000, Synaptic Systems, 251 003), rabbit GFP (1:5000, Synaptic Systems, 132 002), mouse Rab5 (1:1000, Synaptic Systems, 108 111), rabbit Rab6 (1:1000, Synaptic Systems, 273 003), rabbit Lamp1 (1:500, Abcam, ab24170), mouse Mic60 (1:1000, Abcam, ab110329), rabbit Cox4 (1:1000, Synaptic Systems, 298 002), rabbit Cyt C (1:1000, Cell Signaling, Beverly, MA, USA, 11940S), rabbit caspase 9 (1:1000, Abcam, Cambridge, UK, ab185719), mouse Sodium Potassium ATPase, subunit α1 (1:1000, Abcam, Cambridge, UK, ab7671), mouse syntaxin 1 (1:1000, Synaptic Systems, 110 001), mouse VAMP2 (1:10000, Synaptic Systems, 104 211), rabbit phospho-tau (Ser400;Thr403;Ser404) (1:1000, Cell Signaling, Beverly, MA, USA, 11837S).

Techniques: Immunostaining, Control, Expressing

Journal: eLife

Article Title: Increased expression of heme-binding protein 1 early in Alzheimer's disease is linked to neurotoxicity

doi: 10.7554/eLife.47498

Figure Lengend Snippet:

Article Snippet: Antibody , anti-carbonic anhydrase I (Rabbit, polyclonal) , Novus Biologicals , Cat#: NBP1-88191 RRID: AB_11017594 , WB (1:250).

Techniques: Sequencing, Isolation, Transfection, Construct, Plasmid Preparation, Retroviral, Virus, Luciferase, Negative Control, Knock-Out, Recombinant, Concentration Assay, Membrane, Software