rabbit polyclonal anti-lif antibodies  (Proteintech)

Journal: BMC Developmental Biology

Article Title: CDC2/SPDY transiently associates with endoplasmic reticulum exit sites during oocyte maturation

doi: 10.1186/1471-213X-9-8

Figure Lengend Snippet: Listing of markers used to establish the identity of cortical structures in immature oocytes.

Article Snippet: The following antibodies and reagents were used (concentration or dilution and catalog number in brackets): mouse monoclonal anti-CDC2 (1 μg/ml; sc-54) and goat polyclonal anti-SEC23 (2 μg/ml; sc-12107) from Santa Cruz Biotechnologies (Santa Cruz, CA, USA); rabbit polyclonal anti-Calnexin (1:250; SPA-860) from Stressgen Biotechnologies (San Diego, CA, USA); mouse monoclonal anti-PSTAIR (1 μg/ml; ab10345, which is directed against EGVPSTAIREISLLKE, a conserved region in cyclin-dependent kinases (CDKs) [ , ]) and rabbit polyclonal anti-gamma-tubulin (1:1000; ab11321) from Abcam (Cambridge, UK); mouse monoclonal anti-NUP153 (4 μg/ml; MMS-102P) from Covance (Berkeley, CA, USA); mouse monoclonal anti-GM130 (1 μg/ml; G65120) and mouse monoclonal anti-cyclin B1 (5 μg/ml; 554179) from BD Biosciences (San Jose, CA, USA); rabbit polyclonal anti-SPDY (2.2 μg/ml; NB100–2521, directed against a conserved region that is present in both isoforms) from Novus Biologicals (Littleton, CO, USA); Mitotracker Deep Red (200 nM; M22426), goat anti-mouse IgG alexa488, goat anti-rabbit IgG alexa568, and rabbit anti-goat IgG alexa488 (20 μg/ml; A11029, A11036, and A11078, respectively) from Molecular Probes (Eugene, OR, USA); donkey anti-mouse IgG Cy3 (6.25 μg/ml; 715-165-151) from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA, USA); rabbit polyclonal anti-P-GM130 (1:100; detects GM130 phosphorylated on serine 25 [ ]) was a generous gift from Dr. Martin Lowe (University of Manchester, UK).

Techniques: Binding Assay

Journal: BMC Developmental Biology

Article Title: CDC2/SPDY transiently associates with endoplasmic reticulum exit sites during oocyte maturation

doi: 10.1186/1471-213X-9-8

Figure Lengend Snippet: Gamma-tubulin, mitochondria, and NUP153 do not associate with ERES . (A-C) 0 h GV stage oocyte labeled for CDC2 (A, green), DNA (A, blue), and gamma-tubulin (B, red). The CDC2-labeled ERES cluster did not label with gamma-tubulin (C). (D-F) 0 h GV stage oocyte labeled for CDC2 (D, green), DNA (D, blue), and mitochondria, using the mitochondrial marker mitotracker (E, red). Mitotracker did not label CDC2-positive ERES (F). (G-I) Confocal section through the same oocyte as in D-F showing the GV. Note that the much lower staining intensity in G-I compared to D-F is solely the result of a ~65 μm change in depth of imaging, since acquisition settings and image enhancement were identical between these images. (J) 0 h GV stage oocyte labeled for NUP153 (green) and DNA (blue). None of the oocytes examined (0/15) showed a cortical domain. The 3 separate double lines around the GV are the result of small size and shape changes between the 3 consecutive sections used to produce the image. Images are either Z-projections of 3 consecutive sections (A-C, J), or single sections (D-I); scale bars represent 20 μm in C, F, I, and J. Arrows indicate CDC2-labeled ERES, and arrowheads denote the position of the GV.

Article Snippet: The following antibodies and reagents were used (concentration or dilution and catalog number in brackets): mouse monoclonal anti-CDC2 (1 μg/ml; sc-54) and goat polyclonal anti-SEC23 (2 μg/ml; sc-12107) from Santa Cruz Biotechnologies (Santa Cruz, CA, USA); rabbit polyclonal anti-Calnexin (1:250; SPA-860) from Stressgen Biotechnologies (San Diego, CA, USA); mouse monoclonal anti-PSTAIR (1 μg/ml; ab10345, which is directed against EGVPSTAIREISLLKE, a conserved region in cyclin-dependent kinases (CDKs) [ , ]) and rabbit polyclonal anti-gamma-tubulin (1:1000; ab11321) from Abcam (Cambridge, UK); mouse monoclonal anti-NUP153 (4 μg/ml; MMS-102P) from Covance (Berkeley, CA, USA); mouse monoclonal anti-GM130 (1 μg/ml; G65120) and mouse monoclonal anti-cyclin B1 (5 μg/ml; 554179) from BD Biosciences (San Jose, CA, USA); rabbit polyclonal anti-SPDY (2.2 μg/ml; NB100–2521, directed against a conserved region that is present in both isoforms) from Novus Biologicals (Littleton, CO, USA); Mitotracker Deep Red (200 nM; M22426), goat anti-mouse IgG alexa488, goat anti-rabbit IgG alexa568, and rabbit anti-goat IgG alexa488 (20 μg/ml; A11029, A11036, and A11078, respectively) from Molecular Probes (Eugene, OR, USA); donkey anti-mouse IgG Cy3 (6.25 μg/ml; 715-165-151) from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA, USA); rabbit polyclonal anti-P-GM130 (1:100; detects GM130 phosphorylated on serine 25 [ ]) was a generous gift from Dr. Martin Lowe (University of Manchester, UK).

Techniques: Labeling, Marker, Staining, Imaging

Journal: International Journal of Molecular Sciences

Article Title: Expression Pattern of α-Tubulin, Inversin and Its Target Dishevelled-1 and Morphology of Primary Cilia in Normal Human Kidney Development and Diseases

doi: 10.3390/ijms22073500

Figure Lengend Snippet: Toluidine blue staining of semi-thin sections, immunofluorescence staining with α-tubulin and 4’,6-diamidino-2-phenylindole dihydrochloride (DAPI), and transmission electron microscopy of the cortex of healthy 1.5-year-old kidney tissue ( a ) and nephrotic syndrome of the Finnish type (CNF) kidney tissue ( b ). Details of microvilli in healthy kidney tissue at the apical surface are shown as higher magnification inset (row a , electron microscopy). Legend: proximal convoluted tubules (pct), distal convoluted tubules (dct), glomeruli (g), tubules (t), proximal tubules cyst (cy), nucleus (n), epithelial cell (ec), primary cilium (arrows), lobulated glomerulus (arrowhead). Immunofluorescence staining with α-tubulin and DAPI shows dysmorphic primary cilia in multicystic dysplastic kidneys (MCDK) ( c ) and focal segmental glomerulosclerosis (FSGS) ( d ). Electron microscopy of FSGS distal tubular cell with extremely long primary cilium (arrows) at the apical surface ( e ). Nucleus (n), basal body (arrowhead), loss of microvilli on the apical surface (*). Details of primary cilia are shown as higher magnification insets. Magnification toluidine blue sections: ×40, scale bar 100 µm. Magnification immunofluorescence staining: ×100, scale bar 40 µm. Scale bar transmission electron microphotos 2 µm.

Article Snippet: Primary antibodies used were rabbit monoclonal anti-alpha tubulin antibody (dilution 1:1000; ab179484, Abcam, Cambridge, UK), Rabbit Polyclonal Anti-inversin antibody (dilution 1:100; ab65187, Abcam, Cambridge, UK), mouse monoclonal DVL-1 antibody (1:150 dilution; sc-8025, Santa Cruz Biotechnology, Dallas, TX, USA) and rabbit polyclonal anti-gamma tubulin antibody (in order to verify primary cilia-specific staining with alpha-tubulin, 1:500 dilution; ab11321, Abcam, Cambridge, UK).

Techniques: Staining, Immunofluorescence, Transmission Assay, Electron Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Expression Pattern of α-Tubulin, Inversin and Its Target Dishevelled-1 and Morphology of Primary Cilia in Normal Human Kidney Development and Diseases

doi: 10.3390/ijms22073500

Figure Lengend Snippet: Immunofluorescence staining of inversin ( a ) and α-tubulin ( b ) in developing kidney tissues (14th and 15th/16th gestational week (GW)) showing nephrogenic zone with stages of development during the formation of the nephron: metanephric cup (mc), renal vesicle (rv), comma-shaped body (cb), S-shaped body (sb), immature glomerulus (ig). Magnification ×40, scale bar 100 µm.

Article Snippet: Primary antibodies used were rabbit monoclonal anti-alpha tubulin antibody (dilution 1:1000; ab179484, Abcam, Cambridge, UK), Rabbit Polyclonal Anti-inversin antibody (dilution 1:100; ab65187, Abcam, Cambridge, UK), mouse monoclonal DVL-1 antibody (1:150 dilution; sc-8025, Santa Cruz Biotechnology, Dallas, TX, USA) and rabbit polyclonal anti-gamma tubulin antibody (in order to verify primary cilia-specific staining with alpha-tubulin, 1:500 dilution; ab11321, Abcam, Cambridge, UK).

Techniques: Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Expression Pattern of α-Tubulin, Inversin and Its Target Dishevelled-1 and Morphology of Primary Cilia in Normal Human Kidney Development and Diseases

doi: 10.3390/ijms22073500

Figure Lengend Snippet: Immunofluorescence staining of α-tubulin and DAPI in the developing and postnatal human kidney tissues ( a – e ). Kidneys of 14th, 15th/16th, 38th GW ( a – c ); kidneys of 1.5- and 7-year-old children ( d , e ). Positive staining of α-tubulin (arrows) is shown in each structure in the cortex through developmental and postnatal phases ( a – e ), proximal convoluted tubules (pct), distal convoluted tubules (dct) and glomeruli ( g ). Details of primary cilia in pct ( d ) and dct ( a – c , e ) are shown as higher magnification insets (white box). Magnification ×40, scale bar 100 µm. The distribution of α-Tubulin positive cells per structure throughout different stages of development and in the postnatal period is shown in graph ( f ). Graph ( g ) (overall number of protein positive cells in observed structures) shows protein expression related to developmental time (Linear regression) and interrelation of α-Tubulin to DVL-1 through development and maturation (Two-Way ANOVA followed by SIDAK’s posthoc test). Data are shown as mean ± SD.

Article Snippet: Primary antibodies used were rabbit monoclonal anti-alpha tubulin antibody (dilution 1:1000; ab179484, Abcam, Cambridge, UK), Rabbit Polyclonal Anti-inversin antibody (dilution 1:100; ab65187, Abcam, Cambridge, UK), mouse monoclonal DVL-1 antibody (1:150 dilution; sc-8025, Santa Cruz Biotechnology, Dallas, TX, USA) and rabbit polyclonal anti-gamma tubulin antibody (in order to verify primary cilia-specific staining with alpha-tubulin, 1:500 dilution; ab11321, Abcam, Cambridge, UK).

Techniques: Immunofluorescence, Staining, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Expression Pattern of α-Tubulin, Inversin and Its Target Dishevelled-1 and Morphology of Primary Cilia in Normal Human Kidney Development and Diseases

doi: 10.3390/ijms22073500

Figure Lengend Snippet: Semiquantitative staining intensity to α-tubulin, inversin and dishevelled-1 (DVL-1) during kidney developing stages and 1.5- and 7-year-old kidney tissues. In each group, 20 structures of interest were observed. * g—glomeruli, pct—proximal convoluted tubules, dct—distal convoluted tubules, GW—gestational week, y—years of postnatal development.

Article Snippet: Primary antibodies used were rabbit monoclonal anti-alpha tubulin antibody (dilution 1:1000; ab179484, Abcam, Cambridge, UK), Rabbit Polyclonal Anti-inversin antibody (dilution 1:100; ab65187, Abcam, Cambridge, UK), mouse monoclonal DVL-1 antibody (1:150 dilution; sc-8025, Santa Cruz Biotechnology, Dallas, TX, USA) and rabbit polyclonal anti-gamma tubulin antibody (in order to verify primary cilia-specific staining with alpha-tubulin, 1:500 dilution; ab11321, Abcam, Cambridge, UK).

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: Expression Pattern of α-Tubulin, Inversin and Its Target Dishevelled-1 and Morphology of Primary Cilia in Normal Human Kidney Development and Diseases

doi: 10.3390/ijms22073500

Figure Lengend Snippet: Double immunofluorescence staining of inversin (green), DVL-1 (red) and DAPI (blue) in pathological kidney tissues in MCDK ( a ), CNF ( b ) and FSGS ( c ): tubules (t), glomeruli (g), pct cyst (c), the height of pct epithelial cells (*). Structure and cell co-localization of inversin and DVL-1 (arrowheads) are shown in the merged sections with details in higher magnification insets. Negative control stainings are shown as insets on inversin and DVL-1 (a). Magnification ×40, scale bar 100 µm. Relationship of α-tubulin ( d ) and inversin ( e ) to DVL-1 expression in MCDK, FSGS and CNF (two-way ANOVA followed by SIDAK’s post hoc test). The difference in epithelial cell height ( f ) of FSGS, CNF and MCDK pct compared to healthy control (one-way ANOVA followed by Tukey’s post hoc test, n = 50). The difference in overall positive cells percentage of α-tubulin, inversin and DVL-1 in MCDK, CNF and FSGS when compared to healthy control ( g – i ), one-way ANOVA followed by Tukey’s post hoc test). Data are shown as means ± SD. Significant differences are indicated by p -value (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: Primary antibodies used were rabbit monoclonal anti-alpha tubulin antibody (dilution 1:1000; ab179484, Abcam, Cambridge, UK), Rabbit Polyclonal Anti-inversin antibody (dilution 1:100; ab65187, Abcam, Cambridge, UK), mouse monoclonal DVL-1 antibody (1:150 dilution; sc-8025, Santa Cruz Biotechnology, Dallas, TX, USA) and rabbit polyclonal anti-gamma tubulin antibody (in order to verify primary cilia-specific staining with alpha-tubulin, 1:500 dilution; ab11321, Abcam, Cambridge, UK).

Techniques: Double Immunofluorescence Staining, Negative Control, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Expression Pattern of α-Tubulin, Inversin and Its Target Dishevelled-1 and Morphology of Primary Cilia in Normal Human Kidney Development and Diseases

doi: 10.3390/ijms22073500

Figure Lengend Snippet: Double immunofluorescence staining with α-tubulin (red), γ-tubulin (green) and DAPI of the healthy 1.5-year-old kidney tissue ( a ) and in MCDK tissue ( b ). Arrows indicate tubulin. Details of primary cilia are shown as higher magnification insets. Graph showing differences in cilium length ( n = 50, one-way ANOVA followed by Tukey’s post hoc test) between healthy control and pathological kidney tissues ( c ). Data are shown as means ± SD. *** p < 0.001, **** p < 0.0001).

Article Snippet: Primary antibodies used were rabbit monoclonal anti-alpha tubulin antibody (dilution 1:1000; ab179484, Abcam, Cambridge, UK), Rabbit Polyclonal Anti-inversin antibody (dilution 1:100; ab65187, Abcam, Cambridge, UK), mouse monoclonal DVL-1 antibody (1:150 dilution; sc-8025, Santa Cruz Biotechnology, Dallas, TX, USA) and rabbit polyclonal anti-gamma tubulin antibody (in order to verify primary cilia-specific staining with alpha-tubulin, 1:500 dilution; ab11321, Abcam, Cambridge, UK).

Techniques: Double Immunofluorescence Staining