pha e (Vector Laboratories)
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Vector Laboratories
pha e
Pha E, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pha e/product/Vector Laboratories
Average 86 stars, based on 10 article reviews
Price from $9.99 to $1999.99
Pha E, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pha e/product/Vector Laboratories
Average 86 stars, based on 10 article reviews
Price from $9.99 to $1999.99
pha e - by Bioz Stars,
2022-08
86/100 stars
Images
1) Product Images from "Glycophenotyping of osteoarthritic cartilage and chondrocytes by RT-qPCR, mass spectrometry, histochemistry with plant/human lectins and lectin localization with a glycoprotein"
Article Title: Glycophenotyping of osteoarthritic cartilage and chondrocytes by RT-qPCR, mass spectrometry, histochemistry with plant/human lectins and lectin localization with a glycoprotein
Journal: Arthritis Research & Therapy
doi: 10.1186/ar4330
Figure Legend Snippet: Lectin histochemical staining profiles in sections of OA cartilage. (a) Binding of PNA to complex chondrons of a severely degenerated cartilage region could be completely blocked with lactose (inset) ascertaining carbohydrate-specific binding. (b) Omission of the incubation step with biotinylated LEA (first-step reagent) from processing excluded probe-independent signal generation. (c, d) ConA staining: staining pattern of MS ≤4 regions included chondrocytes in deep zones of cartilage (c) . Intense staining of matrix and chondrons (inset) in MS ≥9 regions (d) . (e) PSA staining: positivity of chondrons (inset) and matrix, predominantly in MS ≥9 cartilage. (f, g) PHA-E staining: whereas MS ≤4 regions were negative (f) , MS ≥9 areas (g) presented positive chondrons (inset) and matrix. (h) PHA-L staining: binding sites were restricted to chondrons (insert) and matrix of MS ≥9 cartilage. (i-j) VAA staining: whereas the chondrons of MS ≤4 areas were negative (i) , reactivity was observed both in chondrons (inset) and matrix of MS ≥9 cartilage (j) . (k) LEA staining: reactivity for chondrons (inset) and matrix of MS ≥9 regions. (l) MAA-I staining: reactivity included chondrons (inset) and matrix of MS ≥9 cartilage. (m, n) SNA staining: weak staining of matrix and no staining of chondrons in MS ≤4 cartilage (m) , whereas both chondrons (inset) and matrix were positive in MS ≥9 regions (n) . (o) DBA staining: positivity in chondrons (inset) and matrix of MS ≥9 cartilage. (p, q) PNA staining: positive chondrocytes sparely distributed in the deeper zones of MS ≤4 cartilage ( p ; arrows, inset). In MS ≥9 cartilage (q) , intense matrix staining was observed, whereas chondrons were mostly negative (inset). (r) JAC staining: absent in chondrons (inset), but present in superficial zones of MS ≥9 cartilage. Bars in inserts of d,e,g,l,n,p,q,r: 50μm. Bars in inserts of h , j , k , o : 100μm. MS, Mankin score; OA, osteoarthritis.
Techniques Used: Staining, Binding Assay, Incubation, Mass Spectrometry
2) Product Images from "Sialylated and sulfated N-Glycans in MDCK and engineered MDCK cells for influenza virus studies"
Article Title: Sialylated and sulfated N-Glycans in MDCK and engineered MDCK cells for influenza virus studies
Journal: Scientific Reports
doi: 10.1038/s41598-022-16605-5
Figure Legend Snippet: Lectin and Western blot analyses of total glycoproteins in MDCK, SIAT1, and hCK cells. Total cell lysates of MDCK, SIAT1, and hCK cell lines were treated with PNGase F (PNGaseF), Neuraminidase A (NeuA), or Neuraminidase S (NeuS), and separated by SDS-PAGE. Total extracts of cells at passage 3 (top) and passage 23 (bottom) ( A–F ), plus PNGaseF/neuraminidases mixture (Enzyme mix) control were loaded on the gels. ( A ) Gel stained with Coomassie Brilliant Blue solution. ( B ) Transferred total glycoproteins analyzed by lectin blot using ConA; ( C ) SNA; ( D ) MAL-I; and ( E ) PHA-E. ( F ) Total glycoproteins from cells at passage 3, and 23 were Western blotted with the antibody O6.
Techniques Used: Western Blot, SDS Page, Staining
3) Product Images from "Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology"
Article Title: Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology
Journal: BMC Biotechnology
doi: 10.1186/s12896-014-0101-0
Figure Legend Snippet: hGGT1 lectin blotting confirms differential ALSA binding affinities. Membrane extracts from normal human kidney and liver tissue or Pichia pastoris -expressed hGGT1 were activity-normalized and subjected to immunoprecipitation with a polyclonal anti-hGGT1 large subunit antibody. Equal volumes from each immunoprecipitation eluate were resolved by SDS-PAGE and affinity blotted with anti-hGGT1 ( hGGT1 , top panel ) or the biotinylated lectins, microvirin ( MVN , second panel ), Phaseolus vulgaris Erythroagglutinin ( Pha - E , third panel ), and Datura stramonium lectin ( DSL , bottom panel ). Expanded views of the immunoblots shown here along with the relevant molecular weight markers are included in Additional file 1 .
Techniques Used: Binding Assay, Activity Assay, Immunoprecipitation, SDS Page, Western Blot, Molecular Weight
4) Product Images from "DNA damage is overcome by TRIP13 overexpression during cisplatin nephrotoxicity"
Article Title: DNA damage is overcome by TRIP13 overexpression during cisplatin nephrotoxicity
Journal: JCI Insight
doi: 10.1172/jci.insight.139092
Figure Legend Snippet: Genomic targeting construct of Trip13 overexpression. ( A ) Targeting construct was designed for integration into the ROSA26 region, and the expression cassette was designed with the CAG promoter-floxed stop codon-Trip13 cDNA-T2A-EGFP-polyA. 2A, protease cleavage site ( Trip13 cDNA was FLAG tagged at the 3′ end); CAG, CMV-IE enhancer/chicken β-actin/rabbit β-globin. ( B ) Genomic DNA isolated from mouse pups was PCR analyzed using specific primers to differentiate wild-type, floxed stop, and Cre-containing mice. Wild-type (WT) = 453 bp; floxed stop (flox) = 616 bp; and Cre = 400 bp. M = 1 kb ladder; lanes 1, 4, and 7 = F1/R1 primers (wild-type); 2, 5, and 8 = F2/R1 primers (floxed Trip13); and 3, 6, and 9 = GGT1-Cre primers (Cre); Trip13 st/st , Trip13 Stop/Stop . ( C ) Western blot analysis of FLAG-tagged TRIP13 and GFP protein expression in harvested tissues from Trip13 ΔStop mice. GAPDH was shown as a loading control. K, kidney; Duo, duodenum; Hrt, heart; Br, brain; Lu, lungs; Spl, spleen. ( D – H ) Immunofluorescence was performed on FFPE Trip13 Stop/Stop and Trip13 ΔStop kidneys to detect GFP expression. GFP was detected by Alexa Fluor 555 fluorescence (red color) in ( G ) Trip13 Stop/Stop and ( D , F , and H ) Trip13 ΔStop kidneys. E shows a negative control kidney section incubated without primary GFP antibody (no primary). To determine proximal tubules ( F and G ) or collecting ducts ( H ), Alexa Fluor 488 (green) fluorescence was detected using PVA-E (brush border of proximal tubules) or DBA (collecting ducts). Nuclei were stained with DAPI (blue). Dashed lines in F indicate PVA-E–positive tubules with minimal to undetectable expression of GFP, showing mosaicism of Cre expression. ( I and J ) Immunohistochemical staining for GFP in cisplatin-treated Trip13 ΔStop and Trip13 Stop/Stop mouse lungs. All DAB-stained sections were counterstained with hematoxylin. Scale bar: 200 μm ( D , E , I , and J ), 100 μm ( F – H ). PVA-E, Phaseolus vulgaris erthyroagglutinin; DBA, Dolicus biflorus agglutinin.
Techniques Used: Construct, Over Expression, Expressing, Isolation, Polymerase Chain Reaction, Mouse Assay, Western Blot, Immunofluorescence, Formalin-fixed Paraffin-Embedded, Fluorescence, Negative Control, Incubation, Staining, Immunohistochemistry
5) Product Images from "Shedding of N-acetylglucosaminyltransferase-V is regulated by maturity of cellular N-glycan"
Article Title: Shedding of N-acetylglucosaminyltransferase-V is regulated by maturity of cellular N-glycan
Journal: Communications Biology
doi: 10.1038/s42003-022-03697-y
Figure Legend Snippet: Increased protein level and activity of GnT-V in complex-type N -glycan-deficient cells. a Scheme of biosynthesis pathway of N -glycans. Glycan symbols are according to the standard symbol nomenclature for glycans 93 . Asn: asparagine. b Lectin blotting of proteins from HEK293 cells treated with or without kifunensine (kif). c In vitro enzyme assays of GnTs (left) and FUT8 (right) in HEK293 cells treated with or without kifunensine. For GnTs, GnGnbi-PA was used as an acceptor substrate. For FUT8, GnGnbi-Asn-PNS was used as an acceptor substrate. The product peak for each enzyme reaction was identified from control reactions using each purified recombinant enzyme, as shown in Supplementary Fig. 1 . d Quantification of the activities of GnTs and FUT8 as measured in ( c ). Error bars represent SD ( n = 4). Statistical analysis was by unpaired Student’s t -test. e Proteins from HEK293 cells treated with or without kifunensine were blotted for GnT-V and GAPDH. Before SDS-PAGE, cell lysates were treated with or without PNGase F (PNG-F). The graph shows quantification of the GnT-V signals in western blots. Error bars represent SD ( n = 3). Statistical analysis was by Welch’s t -test. f Proteins from wild-type (WT), GnT-I-deficient (I-KO), and GnT-I (myc-tagged)-rescued HEK293 cells were blotted with Con A, PHA-L4, or anti-myc antibody. g Proteins from WT, GnT-I-deficient (I-KO), and GnT-I-rescued HEK293 cells were treated with or without PNGase F and blotted for GnT-V and GAPDH. The graph shows quantification of the GnT-V signals in PNGase F-treated blots. Error bars represent SD ( n = 3). Statistical analysis was by one-way analysis of variance (ANOVA) with post-hoc Tukey test. h The cellular activity of GnT-V in WT, GnT-I-deficient (I-KO), and GnT-I-rescued HEK293 cells. Error bars represent SD ( n = 3). Statistical analysis was by one-way ANOVA with post-hoc Tukey test. i Localization of Halo-tagged GnT-V (V-Halo) visualized in B16 cells treated with or without kifunensine. (Upper) GM130 and Golgin97 were stained as cis - and trans -Golgi markers, respectively. (Lower) Line plots of relative fluorescence intensity are shown. Arrows indicate the plotted regions. Scale bars: 10 or 2 µm. Staining with Calnexin, an ER marker, was shown in Supplementary Fig. 5 . * p
Techniques Used: Activity Assay, In Vitro, Purification, Recombinant, SDS Page, Western Blot, Staining, Fluorescence, Marker
Figure Legend Snippet: Inhibition of SPPL3-dependent secretion of GnT-V in complex-type N -glycan-deficient cells. a Relative mRNA expression of MGAT5 (encoding GnT-V) in WT, GnT-I-deficient (I-KO), and GnT-I-rescued HEK293 cells. The levels of MGAT5 mRNA were normalized by those of rRNA. Error bars represent SD ( n = 3). Statistical analysis was by one-way ANOVA with post-hoc Tukey test. b Proteins from HEK293 cells treated with dimethylsulfoxide (DMSO), MG132, or chloroquine (CQ) were blotted for GnT-V, β-catenin, p62, or GAPDH. The samples were treated with PNGase F (PNG-F) before SDS-PAGE. c The activity of GnT-V in DMSO-, MG132-, or CQ-treated HEK293 cells. Error bars represent SD ( n = 3). Statistical analysis was by one-way ANOVA with post-hoc Dunnett test. d Proteins in the lysates of WT and GnT-I-deficient cells and in the culture media of these cells were blotted for GnT-V or GAPDH. e WT and SPPL3-deficient B16 cells were treated with or without kifunensine (kif). Proteins in the cell lysates and the culture media of these cells were blotted for GnT-V, APP, or GAPDH. The samples were treated with or without PNGase F before SDS-PAGE. APP was blotted as a positive control of secreted protein 94 . The proteins in the cell lysates were also blotted with Con A or PHA-L4. f The cellular activity of GnT-V in WT and SPPL3-KO B16 cells treated with or without kifunensine. Error bars represent SD ( n = 3). Statistical analysis was by two-way ANOVA with post-hoc Sidak test. *** p
Techniques Used: Inhibition, Expressing, SDS Page, Activity Assay, Positive Control
Figure Legend Snippet: Requirement of glycan terminal modifications for efficient GnT-V shedding. a Proteins from WT and SLC35C1-KO (C1-KO) HEK293 cells were blotted with AAL or anti-actin antibody. b The activity of GnT-V in WT and SLC35C1-KO HEK293 cells. Error bars represent SD ( n = 4). Statistical analysis was by unpaired Student’s t -test. c C -terminally 3 × FLAG-tagged GnT-V WT was expressed in WT and SLC35C1-KO HEK293 cells, and secretion of GnT-V in these cells were assessed by quantify the signal ratio of secreted GnT-V (in medium) to cellular GnT-V. Error bars represent SD ( n = 3). Statistical analysis was by unpaired Student’s t -test. d Proteins from WT, SLC35A2-KO (A2-KO), and SLC35A2-rescued HEK293 cells treated with or without PNGase F were blotted with RCA-I, and anti-GnT-V and anti-GAPDH antibodies. The graph shows quantification of the GnT-V signals in PNGase F-treated blots. Error bars represent SD ( n = 5). Statistical analysis was by one-way ANOVA with post-hoc Tukey test. e The cellular activity of GnT-V. Error bars represent SD ( n = 4). Statistical analysis was by one-way ANOVA with post-hoc Tukey test. f Proteins from WT, SLC35A1-KO (A1-KO), and SLC35A1-rescued HEK293 cells treated with or without PNGase F were blotted with RCA-I, and anti-GnT-V and anti-GAPDH antibodies. The graph shows quantification of the GnT-V signals in PNGase F-treated blots. Error bars represent SD ( n = 4). Statistical analysis was by one-way ANOVA with post-hoc Tukey test. g The cellular activity of GnT-V. Error bars represent SD ( n = 4). Statistical analysis was by one-way ANOVA with post-hoc Tukey test. h Cellular and secreted GnT-V activity from WT, A1-KO and A2-KO HEK293 cells were examined in vitro. The graph shows the relative activity of the ratio of secreted GnT-V (in medium) to cellular GnT-V. Error bars represent SD ( n = 3). Statistical analysis was by one-way ANOVA with post-hoc Dunnett test. i Proteins from WT, GnT-III-KO (III-KO), and GnT-III (myc-tagged)-rescued HEK293 cells treated with or without PNGase F were blotted with PHA-E4, and anti-myc, anti-GnT-V, and anti-GAPDH antibodies. The graph shows quantification of the GnT-V signals in PNGase F-treated blots. Error bars represent SD ( n = 4). Statistical analysis was by one-way ANOVA with post-hoc Tukey test. j The cellular activity of GnT-V. Error bars represent SD ( n = 4). Statistical analysis was by one-way ANOVA with post-hoc Tukey test. k Proteins from WT, GnT-IVa and IVb-DKO (IVa,b-DKO), and GnT-IVa (myc-tagged)-rescued HEK293 cells were blotted with DSA and anti-myc antibody. l The cellular activity of GnT-V. Error bars represent SD ( n = 4). Statistical analysis was by one-way ANOVA with post-hoc Tukey test. m GnT-V WT or its catalytically inactive E297A mutant was expressed in GnT-V-KO (V-KO) HEK293 cells. Proteins in the cell lysates and the culture media of these cells were blotted with PHA-L4, and anti-GnT-V, anti-APP, anti-ZsGreen1, or anti-actin antibodies. The graph shows quantification of the signal ratio of secreted GnT-V to cellular GnT-V. APP was blotted as a positive control of secreted protein. Error bars represent SD ( n = 3). Statistical analysis was by Welch’s t -test. * p
Techniques Used: Activity Assay, In Vitro, Mutagenesis, Positive Control
