Hor 250

Manufactured by ProSpec

The HOR-250 is a precision laboratory instrument designed for conducting advanced chemical and scientific analyses. It features a high-resolution optical readout and a robust, durable construction. The core function of the HOR-250 is to provide accurate and reliable measurement capabilities for a variety of research and testing applications.

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Lab products found in correlation

Hor 272, by ProSpec (4 mentions) H3506, by Merck Group (1 mentions) K rvcl, by Cook Medical (1 mentions) Minc benchtop incubator, by Cook Medical (1 mentions) Hiscribe t7 quick high yield rna synthesis kit, by New England Biolabs (1 mentions) Gene fragments, by Integrated DNA Technologies (1 mentions) M8410, by Merck Group (1 mentions) Mr 051 f, by Merck Group (1 mentions) H4272, by Merck Group (1 mentions)

6 protocols using hor 250

Maximizing Zygote Yield from C57BL/6J Mice

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C57BL/6J (JAX Stock # 000664) donor female mice (age 3 weeks) were superovulated to maximize embryo yield. Each donor female received five International Units (IU) of Pregnant Mare Serum Gonadotropin (PMSG, ProSpec HOR-272) intraperitoneally (IP), followed 47 hours later by 5 IU of human chorionic gonadotropin (hCG, ProSpec HOR-250), IP. Immediately post-administration of hCG, the female was mated with a single C57BL/6J stud male and was checked 22 hours later for the presence of a copulation plug. Females displaying a copulation plug were euthanized and the oviducts excised and placed into M2 medium. Prior to clutch collection the oviducts were placed in M2 medium containing hyaluronidase (Sigma H3506, 0.3 mg/mL). The oocyte clutch was removed by mechanically lysing the ampulla and the clutch was allowed to incubate in the hyaluronidase-containing M2 medium until the cumulus mass had disintegrated to the point of exposing the oocytes/prospective zygotes. The oocytes/prospective zygotes were then transferred through several washes of fresh M2 medium and then, through the process of visual grading, individual identified zygotes were separated and transferred to microdrops of K-RCVL (COOK K-RVCL) medium that were equilibrated under mineral oil (Sigma M8410) for 24 hours in a COOK MINC benchtop incubator (37°C, 5%CO₂/5%O₂/N₂).

Leidy-Davis T., Cheng K., Goodwin L.O., Morgan J.L., Juan W.C., Roca X., Ong S.T, & Bergstrom D.E. (2018). Viable Mice with Extensive Gene Humanization (25-kbp) Created Using Embryonic Stem Cell/Blastocyst and CRISPR/Zygote Injection Approaches. Scientific Reports, 8, 15028.

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Superovulation Protocol for Mouse Breeding

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PMSG (ProSpec, HOR 272) and hCG (ProSpec, HOR 250) were diluted at a dose of 50 IU in sterile water and frozen at −20 °C until further use. Before injection, aliquots were thawed and diluted in sterile water to get a 100 µL dosage of 5 IU for intraperitoneal injection.
PMSG and hCG were given at an interval of 48 h. Hormones were given at 3:00 p.m. at a light cycle of 12:12-h light:dark cycle, lights on at 7:00 a.m. At the time of hCG injection, females were mated with males overnight and checked for plug in the following morning.
For the genetically modified mice at a C57BL/6J background historical data, males used were between 4 and 6 months of age, with proved fertility and donor females between 3 and 5 weeks of age.
For the C57BL/6J experiment, males were between 4 and 6 months of age, with proved fertility; males were used once per week allowing a one week resting period between matings. C57BL/6J donor females were between 3 and 5 weeks of age.
The B6129F1 experiment used 129/SvPasCrl males between 3 and 5 months of age and a similar breeding scheme as the one described for the C57BL/6J males. These males were crossed with C57BL/6J donor females between 3 and 5 weeks of age.

Lamas S., Franquinho F., Morgado M., Mesquita J.R., Gärtner F, & Amorim I. (2020). C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size. Animals : an Open Access Journal from MDPI, 10(8), 1424.

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Hormone-Induced Ovulation in Mice

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CD-1® IGS female mice were injected once on PD6 and additionally on PD13 with saline (n=3) or 4 mg/kg mPD-1 (n=4). On PD18, all mice were injected intraperitoneally (i.p.) with 10 IU pregnant mare serum gonadotropin (PMSG) (HOR-272, ProSpec-Tany TechnoGene Ltd., East Brunswick, NJ) to mimic the oocyte maturation effect of endogenous follicle-stimulating hormone. 48 hours post PMSG injection, all mice were injected i.p. with 5 IU human chorionic gonadotropin (hCG) (HOR-250, ProSpec-Tany TechnoGene Ltd.) to mimic the ovulation induction effect of luteinizing hormone. 14 hours post hCG administration, all mice were euthanized, and all cumulus-oocyte-complexes (COCs) were collected from the oviduct and quantified.

Xu P.C., Luan Y., Yu S.Y., Xu J., Coulter D.W, & Kim S.Y. (2021). Effects of PD-1 blockade on ovarian follicles in a prepubertal female mouse. The Journal of endocrinology, 252(1), 15-30.

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Induction and Assessment of Ovulation

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Ovaries containing preovulatory follicles were obtained by intraperitoneal injection of 22–24-day-old mice with 5 I.U. equine chorionic gonadotropin (eCG) (ProSpec #HOR-272). 44 hours later, the mice were injected intraperitoneally with 1 nmol kisspeptin-54 (Cayman Chemical, #24477) to stimulate an endogenous LH surge [40 (link)]. Ovaries were dissected at indicated time points after kisspeptin injection.
To assess ovulation, cumulus-oocyte complexes were collected from oviducts dissected after hormone injections as described above. For Figure 1B, some mice were injected with human chorionic gonadotropin (hCG, 5 I.U., ProSpec #HOR-250) instead of kisspeptin. Cumulus-oocyte complexes were dissociated by pipetting and then counted.

Owen C.M, & Jaffe L.A. (2023). Luteinizing hormone stimulates ingression of mural granulosa cells within the mouse preovulatory follicle. bioRxiv.

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Generating Knockout Mice with CRISPR-Cas9

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Two sgRNAs were synthesized using the HiScribe Quick T7 High Yield RNA Synthesis Kit (NEB #E2050) following the manufacturer’s instructions. Template DNA used in the reaction were two gBlocks Gene Fragments of 125bp in length obtained from Integrated DNA Technologies. These sgRNAs were designed to target Cas9 to the 5’ UTR and intron 5 of the murine IDS gene (Figure 1). Cas9 encoding mRNA was purchased from TriLink Biotechnologies. sgRNAs and Cas9 mRNA were mixed in RNase-free water. Ten NSG female mice aged between 8 - 9 weeks were super-ovulated by first injection of 5 IU of Pregnant Mare Serum Gonadotropin (PMSG, C1063, Sigma), intraperitoneal (i. p.), per female, and 48 hours later, followed by injection of 5 IU of human Chorionic Gonadotropin (hCG, HOR-250, PROSPEC Protein Specialists), i. p., per female, and these mice were immediately crossed to male NSG mice. The next morning, zygotes were collected and washed using standard methods¹¹. The sgRNA+Cas9 mRNA mixture was injected into the pronuclei of zygotes using standard procedures. The injected embryos were then transferred into pseudo-pregnant CD-1 (Charles River Laboratory) females as previously described [11 ]. Blood or tissues of offspring generated from this process were subsequently genotyped for IDS gene knockout as described below.

Smith M.C., Belur L.R., Karlen A.D., Podetz-Pedersen K., Erlanson O., Laoharawee K., Furcich J., Lund T.C., You Y., Seelig D., Webber B.R, & McIvor R.S. (2023). Generation and characterization of an immunodeficient mouse model of mucopolysaccharidosis type II. Molecular genetics and metabolism, 138(4), 107539.

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Superovulation and Zygote Isolation in C57BL/6J Mice

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24–28-day old female C57BL/6J mice were purchased directly from Jackson Labs (000664) and were superovulated by i.p. injection of 5 IU/mouse of Pregnant Mare Serum Gonadotropin (PMSG, C1063, Sigma). After 47–48 h, 5 IU/mouse of human Chorionic Gonadotropin (hCG, HOR-250, PROSPEC Protein Specialists) was injected i.p. in PMSG-treated females. Superovulating females were immediately crossed with C57BL/6J males at a 1:1 ratio to produce 1-cell zygotes. The next morning, zygotes were collected and washed using standard methods⁷⁰. Briefly, cumulus-oocyte-complex were collected from the ampulla of the plugged females, treated in hyaluronidase (H4272, Sigma) in a 35 mm TC-treated dish (#353001, Falcon) containing 3.5 ml of modified Human Tubal Fluid (mHTF, http://card.medic.kumamoto-u.ac.jp/card/english/sigen/manual/medium/htf.html)^{71 (link)} for 2 min to remove cumulus cells around the zygotes. The zygotes were then washed 2× in mHTF and then zona pellucida was thinned by briefly treating the zygotes in the Acidic Tyrode’s solution (T1788, Sigma). Zygotes were subsequently washed 6X in M2 media (MR-051-F, Millipore), and incubated in 50 μl of mHTF containing rAAV (1.5 × 10⁸ GC/μl) covered by mineral oil (M8410, Sigma) in a 60 mm tissue culture dish (353004, Falcon) for 6 h at a 37 ^oC, 5% CO₂.

Rollins M.R., Raynor J.F., Miller E.A., Butler J.Z., Spartz E.J., Lahr W.S., You Y., Burrack A.L., Moriarity B.S., Webber B.R, & Stromnes I.M. (2023). Germline T cell receptor exchange results in physiological T cell development and function. Nature Communications, 14, 528.

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