Get effective polyclonal antisera in one month

Hu, Yifeng; Guo, Ju Yuan; Shen, Lu; Chen, Yan; Zhang, Zu Chuan; Zhang, Yong Lian

doi:10.1038/sj.cr.7290122

Cited by 63 publications

(44 citation statements)

References 5 publications

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“…With the purified recombinant GST-VCY fusion protein as the antigen, according to a simply modified procedure [7], specific anti-GST-VCY serum was raised in one month.…”

Section: Preparation Of Gst-vcy Polyclone Antibodymentioning

confidence: 99%

“…After being permeated with 0.2% Triton X-100 (in PBS), the fixed cells were incubated in 100 g/ml DNasefree RNase in PBS at 37 o C for 20 min, and washed twice with PBS (pH 7.0). Cells were stained with PI (2 g/ml in PBS) for 5 min in subdued lighting [7]. The coverslips were then soaked in PBS for 5 min followed by at least two rinses.…”

Section: Localization and Colocalization Of Vcx/y Proteins In Cos7 Cellmentioning

confidence: 99%

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Expression and localization of VCX/Y proteins and their possible involvement in regulation of ribosome assembly during spermatogenesis

Zou

Zhang²,

Zhang

et al. 2003

Cell Res

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Variable Charge X/Y (VCX/Y) is a human testis-specific gene family that localized on X and Y chromosomes. In this study, VCY protein was expressed in E. coli in the form of glutathione-S-transferase (GST) fusion protein. With the purified fusion protein as antigen, the anti-GST-VCY antibody was generated and the localization of VCY protein in human testis was determined by immunohistochemistry. In the testis seminiferous epithelium, VCY proteins were highly expressed in nuclei of germ cells. Using propidium iodide staining and green fluorescent protein (GFP) tag technologies, VCY and VCX-8r proteins were mainly localized in the nucleoli of COS7 cells. In addition, the colocalization for VCY and VCX-8r in COS7 cells was also observed. With VCY cDNA as bait, a cDNA fragment of acidic ribosomal protein PO was obtained using yeast two-hybrid system. All the information above indicates that VCX/Y protein family might be involved in the regulation of ribosome assembly during spermatogenesis.

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“…With the purified recombinant GST-VCY fusion protein as the antigen, according to a simply modified procedure [7], specific anti-GST-VCY serum was raised in one month.…”

Section: Preparation Of Gst-vcy Polyclone Antibodymentioning

confidence: 99%

Section: Localization and Colocalization Of Vcx/y Proteins In Cos7 Cellmentioning

confidence: 99%

Expression and localization of VCX/Y proteins and their possible involvement in regulation of ribosome assembly during spermatogenesis

Zou

Zhang²,

Zhang

et al. 2003

Cell Res

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“…The pre-immunized rabbits' blood was obtained, and the serum was separated and kept at K80 8C for later use (negative control). The rabbits were immunized a total of three times with the recombinant ZAG protein and the adjuvants to raise antibodies (Hu et al 2002). The titer of the antiserum was measured using an indirect ELISA at 405 nm using an Anthos Zenyth 1100 multimode detector (Anthos Labtec Instruments GmbH, Wals, Austria) with a 5-s pre-read shake.…”

Section: Production Of Polyclonal Rabbit Anti-zag Antibodiesmentioning

confidence: 99%

The role of Zn-α2 glycoprotein in sperm motility is mediated by changes in cyclic AMP

Qü¹,

Ying²,

Guo³

et al. 2007

Reproduction

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Sperm motility is essential for male reproduction or natural fertilization. The cyclic AMP (cAMP)/cAMP-dependent protein kinase A (PKA) signaling pathway is generally recognized as one of the significant signaling pathways in the regulation of mammalian spermatozoan motility. Since Zn-a2-glycoprotein (ZAG) activity in mammalian adipose tissue is mediated via the b 3 -adrenoreceptor, with upregulation of the cAMP pathway, we hypothesize that ZAG may play the same role in sperm motility regulation, a new factor of regulation of sperm motility. Therefore, the gene encoding human ZAG was cloned and polyclonal antibodies were generated, and then laser scanning confocal microscopy and flow cytometry were employed to identify this protein in human spermatozoa. The results showed that ZAG protein was mostly localized on the pre-equatorial region covering the acrosome, neck, and middle piece of the flagellum of spermatozoa. Furthermore, using computer-assisted sperm analysis, we found that anti-human ZAG antibodies could significantly reduce the motility of human swim-up spermatozoa after 90-or 120-min incubation (P!0.05 and P!0.01 respectively), together with the decreasing of intracellular cAMP and PKA levels. In conclusion, these data suggest that ZAG is present in human spermatozoa and may be involved in the regulation of sperm motility via the cAMP/PKA signaling pathway.

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“…Subsequently, 6-wk-old male New Zealand White rabbits weighing ∼1.5 kg each were immunized with 100 mg purified DrexSIGIRR protein in CFA or IFA four times, followed by collection of anti-serum. The Ab titer was determined by microplate-based ELISA, and specificity to the DrSIGIRR protein was assessed via Western blot analysis (35,36).…”

Section: Preparation Of Recombinant Protein and Polyclonal Abmentioning

confidence: 99%

Characterization of SIGIRR/IL-1R8 Homolog from Zebrafish Provides New Insights into Its Inhibitory Role in Hepatic Inflammation

Feng

Nie

et al. 2016

The Journal of Immunology

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Single Ig IL-1R–related molecule (SIGIRR, also called IL-1R8 or Toll/IL-1R [TIR]8), a negative regulator for Toll/IL-1R signaling, plays critical roles in innate immunity and various diseases in mammals. However, the occurrence of this molecule in ancient vertebrates and its function in liver homeostasis and disorders remain poorly understood. In this study, we identified a SIGIRR homology from zebrafish (Danio rerio [DrSIGIRR]) by using a number of conserved structural and functional hallmarks to its mammalian counterparts. DrSIGIRR was highly expressed in the liver. Ablation of DrSIGIRR by lentivirus-delivered small interfering RNA in the liver significantly enhanced hepatic inflammation in response to polyinosinic-polycytidylic acid [poly(I:C)] stimulation, as shown by the upregulation of inflammatory cytokines and increased histological disorders. In contrast, depletion of TIR domain–containing adaptor inducing IFN-β (TRIF) or administration of TRIF signaling inhibitor extremely abrogated the poly(I:C)-induced hepatic inflammation. Aided by the zebrafish embryo model, overexpression of DrSIGIRR in vivo significantly inhibited the poly(I:C)- and TRIF-induced NF-κB activations; however, knockdown of DrSIGIRR promoted such activations. Furthermore, pull-down and Duolink in situ proximity ligation assay assays showed that DrSIGIRR can interact with the TRIF protein. Results suggest that DrSIGIRR plays an inhibitory role in TRIF-mediated inflammatory reactions by competitive recruitment of the TRIF adaptor protein from its TLR3/TLR22 receptor. To our knowledge, this study is the first to report a functional SIGIRR homolog that existed in a lower vertebrate. This molecule is essential to establish liver homeostasis under inflammatory stimuli. Overall, the results will enrich the current knowledge about SIGIRR-mediated immunity and disorders in the liver.

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Get effective polyclonal antisera in one month

Cited by 63 publications

References 5 publications

Expression and localization of VCX/Y proteins and their possible involvement in regulation of ribosome assembly during spermatogenesis

Expression and localization of VCX/Y proteins and their possible involvement in regulation of ribosome assembly during spermatogenesis

The role of Zn-α2 glycoprotein in sperm motility is mediated by changes in cyclic AMP

Characterization of SIGIRR/IL-1R8 Homolog from Zebrafish Provides New Insights into Its Inhibitory Role in Hepatic Inflammation

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