polyclonal anti-pde4 antibody  (Cusabio)

Journal: Journal of Virology

Article Title: Begomovirus capsid proteins interact with cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase of its whitefly vector and modulate virus retention within its vector

doi: 10.1128/jvi.02172-24

Figure Lengend Snippet: Primers used in this study ^a

Article Snippet: The PDE4 protein expression was measured by western blot as mentioned earlier; 30 μg total protein was loaded in each lane of 12% polyacrylamide gel, and the PDE4 level was detected using 1:5,000 dilution of a polyclonal anti-PDE4 antibody generated by immunizing two rabbits with a 243 amino acid long partial PDE4 protein (NDESVLENHHLVVEFKLLQKEGCDIFINLSKKQKQTLRKMVIDMVLSTDMSKHMSLLADLKAMVETKKVAGSGVLLLDNYTDRIQVLENLVHCADLSNPTKPLPLYRRWVDLLMEEFFQQGDKEREQNLDISPMCDRHSATIEKSQVGFIDYIVHPLWETWADLVHPDAQEILDMLEENRDWYQSMIPPSPPVNEGENRLDSDVEEGEESEPPNPNPPPVPQDSSIRFQVTLEEGDEDSTAQM) expressed in Escherichia coli (CUSABIO, Houston, Texas, USA).

Techniques: Sequencing, Amplification, Cloning

Journal: Journal of Virology

Article Title: Begomovirus capsid proteins interact with cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase of its whitefly vector and modulate virus retention within its vector

doi: 10.1128/jvi.02172-24

Figure Lengend Snippet: Interaction of PDE4 with CP proteins of CuLCrV, SiGMV, TYLCV, and empty pGBKT7 by one-to-one Y2H mating assays on ( A ) stringent −ALTH and ( B ) less stringent −LTH minimal medium supplemented with α-Xgal.

Article Snippet: The PDE4 protein expression was measured by western blot as mentioned earlier; 30 μg total protein was loaded in each lane of 12% polyacrylamide gel, and the PDE4 level was detected using 1:5,000 dilution of a polyclonal anti-PDE4 antibody generated by immunizing two rabbits with a 243 amino acid long partial PDE4 protein (NDESVLENHHLVVEFKLLQKEGCDIFINLSKKQKQTLRKMVIDMVLSTDMSKHMSLLADLKAMVETKKVAGSGVLLLDNYTDRIQVLENLVHCADLSNPTKPLPLYRRWVDLLMEEFFQQGDKEREQNLDISPMCDRHSATIEKSQVGFIDYIVHPLWETWADLVHPDAQEILDMLEENRDWYQSMIPPSPPVNEGENRLDSDVEEGEESEPPNPNPPPVPQDSSIRFQVTLEEGDEDSTAQM) expressed in Escherichia coli (CUSABIO, Houston, Texas, USA).

Techniques:

Journal: Journal of Virology

Article Title: Begomovirus capsid proteins interact with cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase of its whitefly vector and modulate virus retention within its vector

doi: 10.1128/jvi.02172-24

Figure Lengend Snippet: Validation of interactions between begomovirus CPs and PDE4 by pull-down assay of B. tabaci PDE4 protein from bacterial lysates expressing 6x-his-PDE4 by using GST-fused CPs of TYLCV, CuLCrV, and SiGMV as baits. Mixture of bacterial lysates expressing the CP and PDE4 was used as samples, and only PDE4 protein was used as positive control for detection. Monoclonal anti-GST and anti-6x-his antibodies were used to detect CPs and PDE4, respectively. The ponceau-stained membrane shows the protein amounts of the individual lanes.

Article Snippet: The PDE4 protein expression was measured by western blot as mentioned earlier; 30 μg total protein was loaded in each lane of 12% polyacrylamide gel, and the PDE4 level was detected using 1:5,000 dilution of a polyclonal anti-PDE4 antibody generated by immunizing two rabbits with a 243 amino acid long partial PDE4 protein (NDESVLENHHLVVEFKLLQKEGCDIFINLSKKQKQTLRKMVIDMVLSTDMSKHMSLLADLKAMVETKKVAGSGVLLLDNYTDRIQVLENLVHCADLSNPTKPLPLYRRWVDLLMEEFFQQGDKEREQNLDISPMCDRHSATIEKSQVGFIDYIVHPLWETWADLVHPDAQEILDMLEENRDWYQSMIPPSPPVNEGENRLDSDVEEGEESEPPNPNPPPVPQDSSIRFQVTLEEGDEDSTAQM) expressed in Escherichia coli (CUSABIO, Houston, Texas, USA).

Techniques: Biomarker Discovery, Pull Down Assay, Expressing, Positive Control, Staining, Membrane

Journal: Journal of Virology

Article Title: Begomovirus capsid proteins interact with cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase of its whitefly vector and modulate virus retention within its vector

doi: 10.1128/jvi.02172-24

Figure Lengend Snippet: Localization of PDE4 protein and TYLCV in the midgut of ( A ) viruliferous and ( B ) non-viruliferous B. tabaci reared on TYLCV-infected and non-host cotton plants, respectively. TYLCV was detected using polyclonal anti-TYLCV CP antibody and cy5 (green) conjugated secondary antibody. PDE4 was detected using polyclonal anti-PDE4 antibody and cy3 (red) conjugated secondary antibody. The overlay of green and red channels is indicated in yellow, and midgut cell nuclei, stained with DAPI, are indicated in blue. The gut area magnified to 60× and 180× are depicted in white square boxes.

Article Snippet: The PDE4 protein expression was measured by western blot as mentioned earlier; 30 μg total protein was loaded in each lane of 12% polyacrylamide gel, and the PDE4 level was detected using 1:5,000 dilution of a polyclonal anti-PDE4 antibody generated by immunizing two rabbits with a 243 amino acid long partial PDE4 protein (NDESVLENHHLVVEFKLLQKEGCDIFINLSKKQKQTLRKMVIDMVLSTDMSKHMSLLADLKAMVETKKVAGSGVLLLDNYTDRIQVLENLVHCADLSNPTKPLPLYRRWVDLLMEEFFQQGDKEREQNLDISPMCDRHSATIEKSQVGFIDYIVHPLWETWADLVHPDAQEILDMLEENRDWYQSMIPPSPPVNEGENRLDSDVEEGEESEPPNPNPPPVPQDSSIRFQVTLEEGDEDSTAQM) expressed in Escherichia coli (CUSABIO, Houston, Texas, USA).

Techniques: Infection, Staining

Journal: Journal of Virology

Article Title: Begomovirus capsid proteins interact with cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase of its whitefly vector and modulate virus retention within its vector

doi: 10.1128/jvi.02172-24

Figure Lengend Snippet: Relative quantitation of PDE4 mRNA in viruliferous whitefly adults post 12, 24, and 48 hours of AAP on ( A ) CuLCrV- or ( B ) TYLCV-infected squash/tomato plants compared with non-viruliferous whitefly (NV) adults exposed to non-infected plants. ( C ) Abundance of PDE4 protein detected by western blot analysis of soluble proteins extracted from viruliferous whitefly adults post 12 hours of virus acquisition (CuLCrV/TYLCV) compared with non-viruliferous (NV) adults. PDE4 band volumes (intensity) normalized to α-tubulin protein bands of viruliferous (CuLCrV/TYLCV) and NV control samples are depicted in bar graphs. ( D ) Quantitation of cAMP (picomoles per milligram whitefly protein) in viruliferous (post 12 hours of CuLCrV acquisition) and non-viruliferous (NV) whitefly adults.

Article Snippet: The PDE4 protein expression was measured by western blot as mentioned earlier; 30 μg total protein was loaded in each lane of 12% polyacrylamide gel, and the PDE4 level was detected using 1:5,000 dilution of a polyclonal anti-PDE4 antibody generated by immunizing two rabbits with a 243 amino acid long partial PDE4 protein (NDESVLENHHLVVEFKLLQKEGCDIFINLSKKQKQTLRKMVIDMVLSTDMSKHMSLLADLKAMVETKKVAGSGVLLLDNYTDRIQVLENLVHCADLSNPTKPLPLYRRWVDLLMEEFFQQGDKEREQNLDISPMCDRHSATIEKSQVGFIDYIVHPLWETWADLVHPDAQEILDMLEENRDWYQSMIPPSPPVNEGENRLDSDVEEGEESEPPNPNPPPVPQDSSIRFQVTLEEGDEDSTAQM) expressed in Escherichia coli (CUSABIO, Houston, Texas, USA).

Techniques: Quantitation Assay, Infection, Western Blot, Virus, Control

Journal: Journal of Virology

Article Title: Begomovirus capsid proteins interact with cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase of its whitefly vector and modulate virus retention within its vector

doi: 10.1128/jvi.02172-24

Figure Lengend Snippet: ( A ) Quantitation of cAMP (picomoles per milligram whitefly protein) in B. tabaci adults fed for 48 hours on 20% sucrose diet with either 0.8% ethanol (control) or 200 µM rolipram, an inhibitor of PDE4. ( B ) Relative quantitation of CuLCrV DNA and ( C ) TYLCV DNA retained (normalized to the β-tubulin gene of the whitefly) in whitefly adults post virus acquisition (24 hours) from infected squash/tomato plants and overnight gut clearing when pre-fed on 20% sucrose diet (48 hours) with 0.8% ethanol (control) or with 200 µM rolipram.

Article Snippet: The PDE4 protein expression was measured by western blot as mentioned earlier; 30 μg total protein was loaded in each lane of 12% polyacrylamide gel, and the PDE4 level was detected using 1:5,000 dilution of a polyclonal anti-PDE4 antibody generated by immunizing two rabbits with a 243 amino acid long partial PDE4 protein (NDESVLENHHLVVEFKLLQKEGCDIFINLSKKQKQTLRKMVIDMVLSTDMSKHMSLLADLKAMVETKKVAGSGVLLLDNYTDRIQVLENLVHCADLSNPTKPLPLYRRWVDLLMEEFFQQGDKEREQNLDISPMCDRHSATIEKSQVGFIDYIVHPLWETWADLVHPDAQEILDMLEENRDWYQSMIPPSPPVNEGENRLDSDVEEGEESEPPNPNPPPVPQDSSIRFQVTLEEGDEDSTAQM) expressed in Escherichia coli (CUSABIO, Houston, Texas, USA).

Techniques: Quantitation Assay, Control, Virus, Infection

Journal: Journal of Virology

Article Title: Begomovirus capsid proteins interact with cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase of its whitefly vector and modulate virus retention within its vector

doi: 10.1128/jvi.02172-24

Figure Lengend Snippet: Relative quantitation of mRNA (normalized to the β-tubulin gene of the whitefly) of ( A ) PDE4 or ( B ) AC in F1 B. tabaci adults reared on tomato plants transformed with PDE4_TRV2 or AC_TRV2 constructs compared with TRV2 control plants.

Article Snippet: The PDE4 protein expression was measured by western blot as mentioned earlier; 30 μg total protein was loaded in each lane of 12% polyacrylamide gel, and the PDE4 level was detected using 1:5,000 dilution of a polyclonal anti-PDE4 antibody generated by immunizing two rabbits with a 243 amino acid long partial PDE4 protein (NDESVLENHHLVVEFKLLQKEGCDIFINLSKKQKQTLRKMVIDMVLSTDMSKHMSLLADLKAMVETKKVAGSGVLLLDNYTDRIQVLENLVHCADLSNPTKPLPLYRRWVDLLMEEFFQQGDKEREQNLDISPMCDRHSATIEKSQVGFIDYIVHPLWETWADLVHPDAQEILDMLEENRDWYQSMIPPSPPVNEGENRLDSDVEEGEESEPPNPNPPPVPQDSSIRFQVTLEEGDEDSTAQM) expressed in Escherichia coli (CUSABIO, Houston, Texas, USA).

Techniques: Quantitation Assay, Transformation Assay, Construct, Control

Journal: Journal of Virology

Article Title: Begomovirus capsid proteins interact with cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase of its whitefly vector and modulate virus retention within its vector

doi: 10.1128/jvi.02172-24

Figure Lengend Snippet: Relative quantitation of ( A ) CuLCrV and ( B ) TYLCV DNA retained (normalized to the β-tubulin gene of the whitefly) in F1 B. tabaci adults reared on either PDE4_TRV2 or TRV2 plants post 24 hours of virus acquisition from infected plants followed by overnight gut clearing on cotton plants. ( C ) Relative quantitation of CuLCrV DNA retained post 24 hours of virus acquisition and overnight gut clearing in F1 adults reared on AC_TRV2 compared to TRV2 control.

Article Snippet: The PDE4 protein expression was measured by western blot as mentioned earlier; 30 μg total protein was loaded in each lane of 12% polyacrylamide gel, and the PDE4 level was detected using 1:5,000 dilution of a polyclonal anti-PDE4 antibody generated by immunizing two rabbits with a 243 amino acid long partial PDE4 protein (NDESVLENHHLVVEFKLLQKEGCDIFINLSKKQKQTLRKMVIDMVLSTDMSKHMSLLADLKAMVETKKVAGSGVLLLDNYTDRIQVLENLVHCADLSNPTKPLPLYRRWVDLLMEEFFQQGDKEREQNLDISPMCDRHSATIEKSQVGFIDYIVHPLWETWADLVHPDAQEILDMLEENRDWYQSMIPPSPPVNEGENRLDSDVEEGEESEPPNPNPPPVPQDSSIRFQVTLEEGDEDSTAQM) expressed in Escherichia coli (CUSABIO, Houston, Texas, USA).

Techniques: Quantitation Assay, Virus, Infection, Control