Ht25a 1kt

Manufactured by Merck Group

Sourced in Japan

The HT25A-1KT is a laboratory equipment product. It is designed to perform a specific core function. Details about its intended use or application are not available in this factual and unbiased description.

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Lab products found in correlation

Bz 2 analyzer, by Keyence (2 mentions) Bz 9000 microscope, by Keyence (2 mentions) Cm1850 cryostat, by Leica (1 mentions) Anti mouse alexa555 secondary antibody, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Sp5 confocal microscope, by Leica (1 mentions) Caseviewer software, by 3DHISTECH (1 mentions) Panoramic scan 2, by 3DHISTECH (1 mentions) Ab21600, by Abcam (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) D2522, by Merck Group (1 mentions)

7 protocols using ht25a 1kt

Quantifying Intimal Hyperplasia in Mice

Cited 3 times

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Following sacrifice, mice were perfused via a cannula in the left ventricle with phosphate-buffered saline for 5 min, followed by 4% paraformaldehyde for 30 min at 100 cm H₂O. The femoral arteries were embedded in paraffin, cut into 5-μm sections, and prepared for Elastica van Gieson staining. Serial sections of the 1 mm proximal region from the incision site of the wire insertion were evaluated using an Elastica van Gieson stain kit (HT25A-1KT; Sigma-Aldrich, Tokyo, Japan) to visualize the internal elastic lamina, as described previously [14 (link),15 (link)]. Specimens were viewed under a BZ9000 microscope (Keyence, Tokyo, Japan). The intimal and medial areas were measured by computerized morphometry using a BZ-II analyzer (Keyence). Intimal hyperplasia was defined as the formation of a neointimal layer medial to the internal elastic lamina. The medial area represents the area between external and internal elastic laminas. The intima-to-media ratio was calculated as the intimal area divided by the media area, as described previously [[12] (link), [13] (link), [14] (link), [15] (link)].

Takahashi H., Nomiyama T., Terawaki Y., Horikawa T., Kawanami T., Hamaguchi Y., Tanaka T., Motonaga R., Fukuda T., Tanabe M, & Yanase T. (2019). Combined treatment with DPP-4 inhibitor linagliptin and SGLT2 inhibitor empagliflozin attenuates neointima formation after vascular injury in diabetic mice. Biochemistry and Biophysics Reports, 18, 100640.

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Elastin Fiber Analysis of Vaginal Wall

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A small portion of the upper vaginal wall of three cohorts: virgin, para3, and multiparous were placed overnight in 100 ml of phosphate buffered saline with 30 g sucrose. The sample was then placed in Lieca Cryo-Gel (SPI supplies Product 02694-AB) and sectioned on a Leica CM1850 cryostat at 10 μm and 16 μm with no noticeable difference between the two. The samples were then placed on microscope slides and stained using a commercial stain kit for elastin (Sigma Aldrich REF HT25A-1KT) following a protocol adapted from Downing et al.[18 (link)]. The slides were placed in a stain solution containing 20 ml haematoxylin solution, 3 ml of ferric chloride solution, 8ml of Weigert’s iodine solution, and 5 ml of deionized water for 10 min. The slides were then placed in a working ferric chloride solution, gently rinsed with water, placed in 100% ethanol for a few seconds, and again gently rinsed with deionized water. The slides were placed Van Geisen solution for 1.5 min, followed by a gentle rinse with 100% ethanol and placed in xylene for a few seconds. Lastly, cover slips were mounted using Eukitt quick-hardening mounting medium (Sigma-Aldrich REF 03989) and left to dry overnight. Analysis of elastic fiber length and tortuosity was performed using software developed by our group in Matlab, used previously in [18 (link)], and is available upon request.

Dhital B., Downing K.T., Gul-E-Noor F., Landau Y., Rathod P., Hirsch S., Chang E.J, & Boutis G.S. (2019). Alterations of elastin in female reproductive tissues arising from advancing parity. Archives of biochemistry and biophysics, 666, 127-137.

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Aortic Histology in Mouse Models

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We chose aortas from studies of AngII-infused C57BL/6J mice (ascending aortopathy) or Ldlr^−/− mice (descending thoracic or abdominal aortopathies)(n = 1/sex chromosome genotype) that had a primary endpoint measure (internal or external aortic diameters) close to the mean for that group. A pair of 5 μm paraffin embedded aortic sections was placed on each of 5 slides. Three more sets of 5 pairs were placed such that each slide had 4 pairs of sections, each pair from a location on the artery 50 μm distant from the adjacent pairs, with 100 μm discarded between each set of 5 slides from thoracic and abdominal segments, and 60 μm discarded between each set from ascending aortas. A slide from each level was selected and stained with Van Gieson (Sigma HT25A-1KT).

AlSiraj Y., Thatcher S.E., Blalock E., Saintilnord W.N., Daugherty A., Lu H.S., Luo W., Shen Y.H., LeMaire S.A., Arnold A.P, & Cassis L.A. (2020). Monosomy X in Female Mice Influences the Regional Formation and Augments the Severity of Angiotensin II-induced Aortopathies. Arteriosclerosis, thrombosis, and vascular biology, 41(1), 269-283.

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Histological Analysis of Intimal Hyperplasia

Cited 2 times

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Following sacrifice, mice were perfused via a cannula in the left ventricle with phosphate-buffered saline for 5 min, followed by 4% paraformaldehyde for 30 min at 100 cm H₂O. The femoral arteries were embedded in paraffin, cut into 5-μm sections, and prepared for Elastica van Gieson staining. Serial sections were evaluated using an Elastica van Gieson stain kit (HT25A-1KT; Sigma-Aldrich, Tokyo, Japan) to visualize the internal elastic lamina as described previously [5 (link), 6 (link), 7 (link)]. Specimens were viewed under a BZ9000 microscope (Keyence, Tokyo, Japan). The intimal and medial areas were measured by computerized morphometry using a BZ-II analyzer (Keyence). Intimal hyperplasia was defined as described previously [5 (link), 6 (link), 7 (link)]. The medial area represents the area between external and internal elastic laminas. The intima-to-media ratio was calculated as the intimal area divided by the media area as described previously [5 (link), 6 (link), 7 (link)].

Horikawa T., Kawanami T., Hamaguchi Y., Tanaka Y., Kita S., Ryorin R., Takashi Y., Takahashi H., Tanabe M., Yanase T., Kawanami D, & Nomiyama T. (2020). Pemafibrate, a PPAR alpha agonist, attenuates neointima formation after vascular injury in mice fed normal chow and a high-fat diet. Heliyon, 6(11), e05431.

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Immunofluorescence Analysis of BM-MSCs

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BM-MSCs were identified by immunofluorescence staining. The samples were incubated in sodium citrate 10 mM and 0.05% Tween-20, pH 6 at 100 °C for 20 min, rinsed with cold PBS, blocked with 0.2% v/v Triton X-100 and 5% v/v donkey serum in PBS for 1 h at room temperature, and incubated with mouse anti-human mitochondria (dilution 1:1,000, Millipore) overnight at 4°C. Finally, the slides were incubated for 1.5 h in the dark with anti-mouse Alexa555 secondary antibody (dilution 1:1,000, Life Technology). Cell nuclei were stained with DAPI (Invitrogen). Images were obtained using a Leica SP5 confocal microscope.
Elastic fibers of the ELR-based hydrogel were detected with an elastic stain (HT25A-1KT, Sigma) according to the manufacturer’s instructions.

Ibáñez-Fonseca A., Rico A., Preciado S., González-Pérez F., Muntión S., García-Briñón J., García-Macías M.C., Rodríguez-Cabello J.C., Pericacho M., Alonso M, & Sánchez-Guijo F. (2022). Mesenchymal Stromal Cells Combined With Elastin-Like Recombinamers Increase Angiogenesis In Vivo After Hindlimb Ischemia. Frontiers in Bioengineering and Biotechnology, 10, 918602.

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Quantifying Aortic Elastic Fiber Thickness

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The aortic tissue sample was fixed for 72 hours at room temperature and embedded in paraffin. Paraffin sections (5 μm) prepared from the aortic wall were stained with elastin Verhoeff-Van Gieson stain (HT25A-1KT; Sigma-Aldrich, St. Louis, MO) according to the manufacturer's instructions, and each section was imaged under panoramic digital slide scanners (Panoramic SCAN II; 3DHISTECH, Budapest, Hungary). Chromatic analysis of the image was performed with CaseViewer software (version 2.2; 3DHISTECH) to measure the thickness of elastic fiber (50 measuring points) for each specimen. The multiple measured values were averaged as 1 single value for each enrolled patient.

Li F., Li X., Wang Y.T., Yu C.T., Yin G., Chen X.Y., Zhao S., Wang W, & Gao Q. (2020). The Etiological Heterogeneity of Bicuspid Aortopathy between Ascending and Root Morphotype. The heart surgery forum, 23(6).

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Morphological Analysis of Aortic and Carotid Arteries

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The aortic root, aortic arch, brachiocephalic and carotid arteries
were removed en bloc, pinned down to maintain tissue
morphology and fixed in 10% formaldehyde for 48h (n=4 per group). Tissues
were embedded in paraffin and sectioned transversely (5-μm thick).
Verhoeff-Van Gieson elastin staining (HT25A-1KT, Sigma, Dorset, UK) and
Trichrome Stain (Masson) Kit (HT15-1KT, Sigma, Dorset, UK) were used to
investigate vessel wall morphology and elastin and collagen fibres,
respectively. Immunohistochemistry for tropoelastin was performed using an
anti-mouse rabbit polyclonal antibody (1:100, Abcam, ab21600, Cambridge, MA,
USA). Vessel wall area was calculated using the Verhoeff-Van Gieson images
as [adventitia area−the luminal area (mm²)] using ImageJ
(NIH). The immunopositive areas were segmented and expressed as normalized
tropoelastin area (%tropoelastin= tropoelastin immunopositive area/vessel
wall area) × 100. Fluorescent microscopy was performed using a custom
synthesized rhodamine-labeled VVGS peptide derivative (rhod-VVGS). Sections
were incubated with a 200nM solution for 24h at 4°C followed by
nuclear counterstain using Hoechst (ThermoFischer 33342, 1:3000, for 15 min
at room temperature). Slides were shielded from light at all times and
mounted with a Mowiol containing 2.5% 1,4-diazobicyclo-[2.2.2]-octane
(DABCO, Sigma, D2522) medium.

Phinikaridou A., Lacerda S., Lavin B., Andia M.E., Smith A., Saha P, & Botnar R.M. (2018). Tropoelastin: A novel marker for plaque progression and instability. Circulation. Cardiovascular imaging, 11(8), e007303.

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