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Cl8942ap

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CL8942AP is a protein-based product used in various laboratory applications. It serves as a core component in experimental procedures, but a detailed description of its specific function or intended use is not available.

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Lab products found in correlation

Ab192256, by Abcam (2 mentions) Ab9661, by Cell Signaling Technology (2 mentions) Alexa 647, by Thermo Fisher Scientific (2 mentions) D3571, by Thermo Fisher Scientific (2 mentions) Prolong gold antifade, by Thermo Fisher Scientific (2 mentions) Ab15580, by Abcam (2 mentions) H 3300, by Vector Laboratories (2 mentions) α actin fitc acta2, by Merck Group (2 mentions) Irak phosphor t209, by Abcam (2 mentions) Ab6673, by Abcam (2 mentions) Gtx109242, by GeneTex (2 mentions) Cf488a, by Biotium (2 mentions) Pecam 1, by Santa Cruz Biotechnology (2 mentions) Ab15323, by Abcam (2 mentions) Ab28364, by Merck Group (1 mentions) D1306, by Agilent Technologies (1 mentions) Sp5 confocal microscope, by Leica camera (1 mentions) Alexa fluor 647 donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 donkey anti rat igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488, by BioLegend (1 mentions)

5 protocols using cl8942ap

1

Histological Analysis of Explanted Islet Grafts

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Formalin-fixed explanted grafts were embedded in paraffin and sectioned at 5–10 µm thickness. Hematoxylin and eosin (H&E) stained sections were imaged using a Leica optical microscope. For immunofluorescence, cells were stained with DAPI (1:10,000, Invitrogen, D1306) and antibodies specific for insulin (1:100, Dako, cat A0564), glucagon (1:250, Biogenex, cat PU039-UP), Mac-2 (1:100, Cederlane, cat CL8942AP), CD31 (1:20, Abcam, cat AB28364), and α-SMA (1:200, Sigma, cat C6198), and images were acquired using a Leica SP5 confocal microscope. For quantification of islet composition in explanted grafts, the percentage of glucagon+ cells (α cells) and of insulin+ cells (β cells) was quantified in each NC or encapsulated islet using ImageJ software (NIH, Bethesda, MD).
De Toni T., Stock A.A., Devaux F., Gonzalez G.C., Nunez K., Rubanich J.C., Safley S.A., Weber C.J., Ziebarth N.M., Buchwald P, & Tomei A.A. (2022). Parallel Evaluation of Polyethylene Glycol Conformal Coating and Alginate Microencapsulation as Immunoisolation Strategies for Pancreatic Islet Transplantation. Frontiers in Bioengineering and Biotechnology, 10, 886483.
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2

Immunofluorescent Staining of Aortic Root Tissues

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Aortic root tissue sections described in 2.6.4 were stained with immunofluorescent antibodies as described previously [2 (link)]. For apoptosis within plaques, Millipore Sigma In Situ Cell Death Detection Kit TMR red (Cat # 12156792910) was used with kit instructions with the following modifications: After proteinase K incubation, tissues were blocked with 10% horse serum in PBS with 1% BSA and subsequently incubated with anti-MAC2 conjugated with Alexa Fluor 488 (Biolegend 125410)for 2 h. The rest of the kit protocol was followed. Primary antibodies: Rat-anti-Mac2 antibody (1:100) (Accurate Chem. CL8942AP)), Rabbit-anti-smooth muscle heavy chain II (1:400) (Abcam ab53219), and donkey-anti-smooth muscle actin-Cy3 (1:200) (Sigma Aldrich C6198)). Secondary antibodies: Alexa Fluor® 647 donkey anti-rabbit IgG (A31573) and Alexa Fluor® 488 donkey anti-rat IgG (A21208) purchased from Life Technologies.
Blackburn C.M., Schilke R.M., Vozenilek A.E., Chandran S., Bamgbose T.T., Finck B.N, & Woolard M.D. (2021). Myeloid-associated lipin-1 transcriptional co-regulatory activity is atheroprotective. Atherosclerosis, 330, 76-84.
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3

Immunohistochemical Profiling of Immune Cells

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Tissues were fixed in 10% neutral-buffered formalin and embedded in paraffin. Sections were stained with H&E. IHC was performed using anti-F4/80 (Abcam, ab6640), anti-Mac2 (Cedarlane, CL8942AP), anti–CSF-1R (Abcam, ab215441), anti-CD45R (BD Biosciences, 550286), anti-MPO (Abcam, ab9535), and anti-EPX (Biorbyt, orb5168) antibodies, and the antigen-antibody binding was detected using Simple Stain MAX PO (Nichirei) and the DAB Substrate Kit (Nichirei).
Matsumoto Y., Dimitriou I.D., La Rose J., Lim M., Camilleri S., Law N., Adissu H.A., Tong J., Moran M.F., Chruscinski A., He F., Asano Y., Katsuyama T., Sada K.E., Wada J, & Rottapel R. (2022). Tankyrase represses autoinflammation through the attenuation of TLR2 signaling. The Journal of Clinical Investigation, 132(7), e140869.
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4

Multiparametric Immunohistochemistry Analysis

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BCA sections were de-paraffinized and rehydrated in xylene and ethanol series. After antigen retrieval (H-3300, Vector Laboratories), sections were blocked with fish skin gelatin–PBS (6 g/L) containing 10% horse serum for 1 hour at room temperature. Slides were incubated with the following antibodies: mouse monoclonal SM α-actin-FITC (ACTA2) (4.4 μg/mL, clone 1A4, Sigma Aldrich), goat polyclonal anti-GFP (4 μg/mL, ab6673, Abcam) for detection of YFP, LGALS3 (2 μg/mL, Cedarlane CL8942AP), RUNX2 (1.374 μg/mL, ab192256, Abcam), Ki67 (4 μg/mL, ab15580, Abcam), PECAM-1 (1 μg/mL, Santa Cruz), IRAK phosphor-T209 (4 μg/mL, Abcam), iNOS (0.52 μg/mL, ab15323, Abcam), and Arg1 (9.2 μg/mL, GTX109242, GeneTex). The secondary antibodies were donkey anti-rat conjugated to Alexa 555 (5 μg/mL, Abcam), donkey anti-rat conjugated to Alexa 647 (5 μg/mL, Abcam), donkey anti-goat conjugated to Alexa 555 (5 μg/mL, Invitrogen), and donkey anti-goat conjugated to Alexa 647 (4 μg/mL, Invitrogen). Apoptosis was assessed by TUNEL (CF488A, Biotium) and cleaved caspase 3 staining (0.84 ug/mL, ab9661, Cell Signaling Technology). IL-1β antibody was detected using a donkey anti-mouse IgG2a-Alexa 488 antibody. DAPI (0.05 mg/mL, D3571, ThermoFisher Scientific) was used as a nuclear counterstain and slides were mounted using Prolong Gold Antifade (Invitrogen).
Gomez D., Baylis R.A., Durgin B.G., Newman A.A., Alencar G.F., Mahan S., St. Hilaire C., Muller W., Waisman A., Francis S.E., Pinteaux E., Randolph G.J., Gram H, & Owens G.K. (2018). Interleukin-1β promotes atheroprotective changes of advanced atherosclerotic lesions in mice. Nature medicine, 24(9), 1418-1429.
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5

Multiparametric Immunohistochemistry Analysis

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BCA sections were de-paraffinized and rehydrated in xylene and ethanol series. After antigen retrieval (H-3300, Vector Laboratories), sections were blocked with fish skin gelatin–PBS (6 g/L) containing 10% horse serum for 1 hour at room temperature. Slides were incubated with the following antibodies: mouse monoclonal SM α-actin-FITC (ACTA2) (4.4 μg/mL, clone 1A4, Sigma Aldrich), goat polyclonal anti-GFP (4 μg/mL, ab6673, Abcam) for detection of YFP, LGALS3 (2 μg/mL, Cedarlane CL8942AP), RUNX2 (1.374 μg/mL, ab192256, Abcam), Ki67 (4 μg/mL, ab15580, Abcam), PECAM-1 (1 μg/mL, Santa Cruz), IRAK phosphor-T209 (4 μg/mL, Abcam), iNOS (0.52 μg/mL, ab15323, Abcam), and Arg1 (9.2 μg/mL, GTX109242, GeneTex). The secondary antibodies were donkey anti-rat conjugated to Alexa 555 (5 μg/mL, Abcam), donkey anti-rat conjugated to Alexa 647 (5 μg/mL, Abcam), donkey anti-goat conjugated to Alexa 555 (5 μg/mL, Invitrogen), and donkey anti-goat conjugated to Alexa 647 (4 μg/mL, Invitrogen). Apoptosis was assessed by TUNEL (CF488A, Biotium) and cleaved caspase 3 staining (0.84 ug/mL, ab9661, Cell Signaling Technology). IL-1β antibody was detected using a donkey anti-mouse IgG2a-Alexa 488 antibody. DAPI (0.05 mg/mL, D3571, ThermoFisher Scientific) was used as a nuclear counterstain and slides were mounted using Prolong Gold Antifade (Invitrogen).
Gomez D., Baylis R.A., Durgin B.G., Newman A.A., Alencar G.F., Mahan S., St. Hilaire C., Muller W., Waisman A., Francis S.E., Pinteaux E., Randolph G.J., Gram H, & Owens G.K. (2018). Interleukin-1β promotes atheroprotective changes of advanced atherosclerotic lesions in mice. Nature medicine, 24(9), 1418-1429.
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