Ab10558

Manufactured by Agilent Technologies

The AB10558 is a laboratory equipment product manufactured by Agilent Technologies. It is designed to perform specific tasks in a research or analytical setting. The core function of this equipment is to provide accurate and reliable measurements, but a detailed description of its intended use or capabilities cannot be provided in an unbiased and factual manner within the given constraints.

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Lab products found in correlation

Ab11089, by Abcam (1 mentions) Alexa fluor 568 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Cm3050, by Leica camera (1 mentions) Mab5428, by Merck Group (1 mentions) Z0334, by BD (1 mentions) Ab28364, by Agilent Technologies (1 mentions) St5020 autostainer, by Leica camera (1 mentions) Envision system hrp dab kit, by Agilent Technologies (1 mentions) Autostainer link 48, by Agilent Technologies (1 mentions) Ab5694, by Abcam (1 mentions) Glial fibrillary acidic protein (gfap), by Agilent Technologies (1 mentions) Ab27957, by Abcam (1 mentions) M0851, by Agilent Technologies (1 mentions) Alexa 647 donkey anti rabbit secondary antibodies, by Jackson ImmunoResearch (1 mentions) Ab13970, by Abcam (1 mentions)

4 protocols using ab10558

Immunohistochemical Analysis of Enucleated Eyes

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Eyes were enucleated and placed in 4% paraformaldehyde (PFA) for less than 30 min, followed by gentle removal of anterior segments, including lens and vitreous. Whole eye cups were fixed in 4% PFA for 2 h at room temperature, rinsed with phosphate-buffered saline (PBS), cryoprotected with 30% sucrose overnight, embedded in optimal cutting temperature (OCT) medium, and then cryosectioned (18 μm) using a cryostat (CM3050 S, Leica). For immunohistochemistry, sections were washed with PBS, blocked with 10% normal donkey serum for 30 min, then incubated in primary antibody for 1–2 h at room temperature, followed by Alexa Fluor 568-conjugated secondary antibodies (Invitrogen). Primary antibodies include RPE65 (MAB5428, 1:250, Millipore), EGFP (Novousbio, NB100-1770, 1:500), IBA-1 (Wako, AB10558, 1:100), GFAP (Dako, Z0334, 1:200), CD45 (BD, 552566, 2.5 μg/mL), and CD3 (Abcam, ab11089, 1:100).

Yiu G., Chung S.H., Mollhoff I.N., Nguyen U.T., Thomasy S.M., Yoo J., Taraborelli D, & Noronha G. (2020). Suprachoroidal and Subretinal Injections of AAV Using Transscleral Microneedles for Retinal Gene Delivery in Nonhuman Primates. Molecular Therapy. Methods & Clinical Development, 16, 179-191.

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Tissue Staining and Immunohistochemistry

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Tissues were dehydrated, paraffin-embedded, and cut into 5-μM thick sections. Trichrome staining (38016SS2; Leica Biosystems, Buffalo Grove, IL) and Herovici staining (American MasterTech Scientific Laboratory Supplies KTHERPT, Lodi, CA) were performed with a Leica ST5020 Autostainer (Richmond, IL). A standard immunohistochemistry (IHC) protocol was followed for all stains, which were performed on a Dako Autostainer Link 48 and its accompanying software (DakoLink version 4.1, edition 3.1.0.987; Agilent, Santa Clara, CA). Primary antibodies—α-smooth muscle actin (α-SMA) for myofibroblasts (ab5694; Abcam, Cambridge, MA), CD45 for pan-leukocytes (ab10558), CD31 for microvasculature (ab28364), and F4/80 for pan-macrophages (ab111101) were detected by EnVision+System-HRP (DAB) kits (Dako North America, Carpinteria, CA).

Wang X., Balaji S., Steen E.H., Li H., Rae M.M., Blum A.J., Miao Q., Butte M.J., Bollyky P.L, & Keswani S.G. (2019). T Lymphocytes Attenuate Dermal Scarring by Regulating Inflammation, Neovascularization, and Extracellular Matrix Remodeling. Advances in Wound Care, 8(11), 527-537.

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Immunohistochemical Analysis of Retinal Cells

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Immunohistochemistry was performed as described previously. 21 (link) Posterior eye cups were fixed with 4% paraformaldehyde (PFA) for 2 hours after removal of anterior segments lens and vitreous. After washing with PBS, tissues were cryoprotected with 30% sucrose overnight, then embedded and cryosectioned at 18µm. For antibody labelling, sections were washed with PBS, blocked with 10% normal donkey serum for 30 min, then incubated in primary antibody for 1-2 hours at room temperature, followed by Alexa Fluor-conjugated secondary antibodies. Primary antibodies include IBA-1 (Wako, AB10558, 1:100), GFAP (Dako, Z0334, 1:200), and CD45 (BD, 552566, 2.5µg/ml).

Chung S.H., Mollhoff I.N., Mishra A., Sin T., Ngo T., Ciulla T.A., Sieving P.A., Thomasy S.M., & Yiu G. (2020). Host immune responses after suprachoroidal delivery of AAV8 in nonhuman primate eyes.

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Histopathological Lung Tissue Analysis

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Tissue samples of control lungs and tumor-bearing lungs were routinely fixed with formalin and embedded in paraffin. Tissues were then serially sectioned in 4 to 7 μM distances and transferred to slides for hematoxylin and eosin staining.
For immuno-fluorescent staining, the following antibodies were used: GFP was amplified by chicken polyclonal antibodies to GFP (ab13970; Abcam, Cambridge, England, United Kingdom) followed by Alexa 488 donkey anti-chicken secondary antibodies (Jackson, 703-545-155). Rabbit pAb to S100A4 (ab27957; Abcam) followed by Alexa 647 donkey anti-rabbit secondary antibodies (Jackson, 711-605-152; JAX Mice, Clinical & Research Services, Bar Harbor, ME) and PE-conjugated anti-mCD45 (clone 30-F11, Biolegend, San Diego, CA). For immunohistochemistry, slides were incubated with anti-CD45 (Abcam, ab10558) or anti-aSMA (DAKO M0851). 3-Amino-9-ethylcarbazole was used for color development and sections were counterstained with hematoxylin.

Weiss I.D., Ella E., Dominsky O., Smith Y., Abraham M., Wald H., Shlomai Z., Zamir G., Feigelson S.W., Shezen E., Bar-Shai A., Alon R., Izhar U., Peled A., Shapira O.M, & Wald O. (2015). In the hunt for therapeutic targets: mimicking the growth, metastasis, and stromal associations of early-stage lung cancer using a novel orthotopic animal model. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 10(1).

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