Generating a Battery of Monoclonal Antibodies Against Native Green Fluorescent Protein for Immunostaining, FACS, IP, and ChIP Using a Unique Adjuvant

Sanchez, Philip; Daniels, Karla J.; Park, Yang-Nim; Soll, David R.

doi:10.1089/mab.2013.0089

Cited by 20 publications

(19 citation statements)

References 33 publications

(47 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…[DHSB-4C9]) (Sanchez et al 2014). ChIPs for Rhino and Cutoff were performed with anti-GFP antibodies on ovaries from transgenic animals expressing GFP-tagged proteins.…”

Section: Chip and Chip-qpcrmentioning

confidence: 99%

Transgenerationally inherited piRNAs trigger piRNA biogenesis by changing the chromatin of piRNA clusters and inducing precursor processing

et al. 2014

View full text Add to dashboard Cite

Small noncoding RNAs that associate with Piwi proteins, called piRNAs, serve as guides for repression of diverse transposable elements in germ cells of metazoa. In Drosophila, the genomic regions that give rise to piRNAs, the socalled piRNA clusters, are transcribed to generate long precursor molecules that are processed into mature piRNAs. How genomic regions that give rise to piRNA precursor transcripts are differentiated from the rest of the genome and how these transcripts are specifically channeled into the piRNA biogenesis pathway are not known. We found that transgenerationally inherited piRNAs provide the critical trigger for piRNA production from homologous genomic regions in the next generation by two different mechanisms. First, inherited piRNAs enhance processing of homologous transcripts into mature piRNAs by initiating the ping-pong cycle in the cytoplasm. Second, inherited piRNAs induce installment of the histone 3 Lys9 trimethylation (H3K9me3) mark on genomic piRNA cluster sequences. The heterochromatin protein 1 (HP1) homolog Rhino binds to the H3K9me3 mark through its chromodomain and is enriched over piRNA clusters. Rhino recruits the piRNA biogenesis factor Cutoff to piRNA clusters and is required for efficient transcription of piRNA precursors. We propose that transgenerationally inherited piRNAs act as an epigenetic memory for identification of substrates for piRNA biogenesis on two levels: by inducing a permissive chromatin environment for piRNA precursor synthesis and by enhancing processing of these precursors.

show abstract

“…[DHSB-4C9]) (Sanchez et al 2014). ChIPs for Rhino and Cutoff were performed with anti-GFP antibodies on ovaries from transgenic animals expressing GFP-tagged proteins.…”

Section: Chip and Chip-qpcrmentioning

confidence: 99%

Transgenerationally inherited piRNAs trigger piRNA biogenesis by changing the chromatin of piRNA clusters and inducing precursor processing

et al. 2014

View full text Add to dashboard Cite

show abstract

“…Cells were then fixed in 4% paraformaldehyde in PBS and stained with the anti-GFP antibody DSHB-GFP-12E6 (Developmental Studies Hybridoma Bank, http://dshb. biology.uiowa.edu, Iowa City, IA) followed by counterstaining with Alexa 488 goat anti-mouse IgG (Life Sciences, Carlsbad, CA) as described in great detail in Sanchez et al (2014). Coverslips were examined with the Bio-Rad Radiance 2100MP confocal microscope and digital images acquired with identical settings on a Canon EOS RebelT3i/EOS 600D SLR camera.…”

Section: Nhe1-gfp Localizationmentioning

confidence: 99%

Huntingtin regulates Ca2+ chemotaxis and K+-facilitated cAMP chemotaxis, in conjunction with the monovalent cation/H+ exchanger Nhe1, in a model developmental system: Insights into its possible role in Huntington׳s disease

Wessels

Lusche

Scherer

et al. 2014

Developmental Biology

Self Cite

View full text Add to dashboard Cite

Huntington׳s disease is a neurodegenerative disorder, attributable to an expanded trinucleotide repeat in the coding region of the human HTT gene, which encodes the protein huntingtin. These mutations lead to huntingtin fragment inclusions in the striatum of the brain. However, the exact function of normal huntingtin and the defect causing the disease remain obscure. Because there are indications that huntingtin plays a role in Ca(2+) homeostasis, we studied the deletion mutant of the HTT ortholog in the model developmental system Dictyostelium discoideum, in which Ca(2+) plays a role in receptor-regulated behavior related to the aggregation process that leads to multicellular morphogenesis. The D. discoideum htt(-)-mutant failed to undergo both K(+)-facilitated chemotaxis in spatial gradients of the major chemoattractant cAMP, and chemotaxis up a spatial gradient of Ca(2+), but behaved normally in Ca(2+)-facilitated cAMP chemotaxis and Ca(2+)-dependent flow-directed motility. This was the same phenotypic profile of the null mutant of Nhel, a monovalent cation/H(+)exchanger. The htt(-)-mutant also failed to orient correctly during natural aggregation, as was the case for the Nhel mutant. Moreover, in a K(+)-based buffer the normal localization of actin was similarly defective in both htt(-) and nhe1(-) cells in a K(+)-based buffer, and the normal localization of Nhe1 was disrupted in the htt(-) mutant. These observations demonstrate that Htt and Nhel play roles in the same specific cation-facilitated behaviors and that Nhel localization is directly or indirectly regulated by Htt. Similar cation-dependent behaviors and a similar relationship between Htt and Nhe1 have not been reported for mammalian neurons and deserves investigation, especially as it may relate to Huntington׳s disease.

show abstract

“…Two 8-week-old female BALB/c mice were immunized by intraperitoneal injection of 50 mg of purified recombinant 6xHis-MBP, combined with 50 mg of poly(I:C) HMW VacciGrade (InvivoGen, San Diego, CA) and 50 mg of anti-CD40 mAb (BioXCell, West Lebanon, NH) as the adjuvant, (13,14) in 200 mL of phosphate buffer solution (PBS; Gibco, Grand Island, NY). Mice were boosted 2 weeks after initial immunization by injection of 50 mg of recombinant 6xHis-MBP and 50 mg of poly(I:C) in 200 mL of PBS.…”

Section: Mice and Immunizationmentioning

confidence: 99%

“…The hybridoma fusion was performed as described previously (14) with minor modification. In brief, spleen cells of the immunized mouse and Sp2/0-Ag14 myeloma cells (CRL-1581; ATCC, Manassas, VA) (15) were washed, passed through a 70 mm filter, and resuspended in serum-free RPMI 1640 medium (HyClone, Logan, Utah).…”

Section: Hybridoma Fusionmentioning

confidence: 99%

See 1 more Smart Citation

Generation and Validation of Monoclonal Antibodies Against the Maltose Binding Protein

Park

Glover

Daniels

et al. 2016

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

Self Cite

View full text Add to dashboard Cite

Generating a Battery of Monoclonal Antibodies Against Native Green Fluorescent Protein for Immunostaining, FACS, IP, and ChIP Using a Unique Adjuvant

Cited by 20 publications

References 33 publications

Transgenerationally inherited piRNAs trigger piRNA biogenesis by changing the chromatin of piRNA clusters and inducing precursor processing

Transgenerationally inherited piRNAs trigger piRNA biogenesis by changing the chromatin of piRNA clusters and inducing precursor processing

Huntingtin regulates Ca2+ chemotaxis and K+-facilitated cAMP chemotaxis, in conjunction with the monovalent cation/H+ exchanger Nhe1, in a model developmental system: Insights into its possible role in Huntington׳s disease

Generation and Validation of Monoclonal Antibodies Against the Maltose Binding Protein

Contact Info

Product

Resources

About