Ab21600

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab21600 is a lab equipment product produced by Abcam. It is a device designed for laboratory use, but a detailed description of its core function cannot be provided in an unbiased and factual manner without potential extrapolation.

Automatically generated - may contain errors

Lab products found in correlation

Duolink proximity ligation assay kit, by Thermo Fisher Scientific (1 mentions) Vectashield, by Vector Laboratories (1 mentions) Ab64409, by Abcam (1 mentions) Ab150077, by Abcam (1 mentions) Sc 398631, by Santa Cruz Biotechnology (1 mentions) Victor nivo, by PerkinElmer (1 mentions) Sc 166572, by Santa Cruz Biotechnology (1 mentions) Ab133642, by Abcam (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) D2522, by Merck Group (1 mentions) Ht25a 1kt, by Merck Group (1 mentions)

8 protocols using ab21600

Histological Evaluation of Atherosclerotic Plaques

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After the imaging experiments, the plaques from endarterectomy were
fixed in formalin, decalcified, embedded in paraffin, and sectioned
transversely at 5μm. Sections were stained for collagen using
Masson’s trichrome, elastin with Verhoeff-Van Gieson and tropoelastin
by immunohistochemistry (1:100, Abcam, ab21600, Cambridge, MA, USA).
Western Blot experiments for quantification of vessel
wall tropoelastin concentration are described in the online Supplementary
Methods.
Inductively coupled plasma mass spectrometry
(ICP–MS) was used to quantify vessel wall gadolinium
concentrations as described in the online Supplementary Methods.

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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Aortic Tissue Morphology Analysis

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The aortic root, aortic arch, and brachiocephalic
and carotid arteries of diseased and control mice were removed en bloc, pinned down to maintain the tissue morphology,
and fixed in 10% formaldehyde for 48 h at 4 °C (n = 3). Tissues were embedded in paraffin and sectioned transversely
(5 μm thick). Immunohistochemistry for tropoelastin was performed
using an anti-mouse rabbit polyclonal antibody (1:100, Abcam, ab21600,
Cambridge, MA, USA).

Capuana F., Phinikaridou A., Stefania R., Padovan S., Lavin B., Lacerda S., Almouazen E., Chevalier Y., Heinrich-Balard L., Botnar R.M., Aime S, & Digilio G. (2021). Imaging of Dysfunctional Elastogenesis in Atherosclerosis Using an Improved Gadolinium-Based Tetrameric MRI Probe Targeted to Tropoelastin. Journal of Medicinal Chemistry, 64(20), 15250-15261.

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Immunohistochemistry and Proximity Ligation for CELA1

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For immunohistochemistry, the ABC Vectastain kit was used with a previously validated anti-CELA1 guinea pig polyclonal antibody (5 (link)). For immunofluorescence imaging, the KF4 anti-CELA1 antibody was used with Abcam anti-tropoelastin antibody (ab21600), both diluted 1:200. For proximity ligation assay, the Duolink Proximity Ligation Assay kit (Thermo Fisher Scientific) was used with these same 2 antibodies.

Ojha M., Smith N.J., Devine A.J., Joshi R., Goodman E.M., Fan Q., Schuman R., Porollo A., Wells J.M., Tiwary E., Batie M.R., Gray J., Deshmukh H., Borchers M.T., Ammerman S.A, & Varisco B.M. (2024). Anti-CELA1 antibody KF4 prevents emphysema by inhibiting stretch-mediated remodeling. JCI Insight, 9(1), e169189.

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Immunohistochemical Analysis of Tropoelastin

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Cryostat sections (20 mm thick) of postnatal lungs were warmed and blocked with 10% goat serum in PBS. The slides were treated with anti-tropoelastin antibody (ab21600, Abcam, Cambridge, MA). The slides were washed twice and treated with secondary Texas-Red-conjugated goat anti-rabbit antibody (T2767, Life Technologies, Carlsbad, CA). After 15-min incubation, the slides were washed twice, mounted in Vectashield (Vector Laboratories, Burlingame, CA) and imaged for analysis.

VALENZUELA C.D., WAGNER W.L., BENNETT R.D., YSASI A.B., BELLE J.M., MOLTER K., STRAUB B., WANG D., CHEN Z., ACKERMANN M., TSUDA A, & MENTZER S.J. (2017). Extracellular Assembly of the Elastin Cable Line Element in the Developing Lung. Anatomical record (Hoboken, N.J. : 2007), 300(9), 1670-1679.

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Immunofluorescent Detection of Procollagen and Elastin

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Five-micron skin sections were stained with rat anti-procollagen type I (#ab64409, Abcam, Cambridge, UK) and rabbit anti-tropoelastin (#ab21600, Abcam, Cambridge, UK) antibodies overnight at 4 °C. Slides were washed and then incubated with secondary antibodies (goat anti-rat Alexa 546 [#A-11081, Invitrogen, Carlsbad, CA] or goat anti-rabbit Alexa 488 [#ab150077, Abcam, Cambridge, UK]) for 1 h at room temperature and counterstaining of cell nuclei was performed using DAPI. For all experimental conditions, a control incubated without primary antibodies was included.

Narda M., Brown A., Muscatelli-Groux B., Grimaud J.A, & Granger C. (2020). Epidermal and Dermal Hallmarks of Photoaging are Prevented by Treatment with Night Serum Containing Melatonin, Bakuchiol, and Ascorbyl Tetraisopalmitate: In Vitro and Ex Vivo Studies. Dermatology and Therapy, 10(1), 191-202.

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ELISA Quantification of Fibrotic Markers

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Per well, 8 × 10³ HDF were grown in 96-well plates and treated with the 0.002% of ObHEx or TGFβ 2.5 ng/mL. After 24 h, the cells were processed for ELISA using monoclonal primary antibody anti-procollagen type I (sc-166572, Santa-Cruz Biotechnology, Dallas, TX, USA), followed by incubation with the secondary anti-mouse antibody labeled with peroxidase (170-6516, Biorad, Hercules, CA, USA). The supernatants of the cells were coated on another plate for the detection of tropoelastin and periostin using the anti-tropoelastin rabbit antibody (ab21600, Abcam, Cambridge, UK) or anti-periostin mouse antibody (sc-398631, Santa-Cruz Biotechnology, Dallas, TX, USA), followed by incubation with secondary antibody anti-rabbit labeled with peroxidase (170-6515, Biorad, Hercules, CA, USA). The colorimetric reaction was developed by adding 100 μL of an aqueous solution of OPD (O-phenylenediamine), 0.35 mg/mL in 50 mM citrate buffer, and 0.012% hydrogen peroxide (H₂O₂). After 30 min, the absorbance was measured at 490 nm using the multiplate reader Victor Nivo (PerkinElmer, Waltham, MA, USA).

Ceccacci S., De Lucia A., Tito A., Tortora A., Falanga D., Arciello S., Ausanio G., Di Cicco C., Monti M.C, & Apone F. (2021). An Oenothera biennis Cell Cultures Extract Endowed with Skin Anti-Ageing Activity Improves Cell Mechanical Properties. Metabolites, 11(8), 527.

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Immunoprecipitation of Lung Proteins

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Ten milligrams of lung homogenate from 2 organ donors was incubated with 100 μg KF4 antibody and KF4 precipitated using Protein A/G beads, washed, and eluting using loading dye. Western blotting was performed using KF4 antibody, anti-AAT antibody (Abcam, ab133642), and anti-tropoelastin antibody (Abcam, ab21600). Revert total protein stain was used for normalization (LICOR, 926-11011). See complete unedited blots in the supplemental material.

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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.

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