Primary antibodies: rat anti-CD31 (DM3614P, Dianova, Hamburg, Germany), goat anti-CD32b (AF1460, R&D Systems, Minneapolis, MN, USA), goat anti-Lama4 (AF3837, R&D Systems, Minneapolis, MN, USA), rabbit anti-Desmin (ab32362, Abcam, Cambridge, UK), rabbit anti-CD3 (100202, Biolegend, San Diego, CA, USA), rat anti-CD4 (100402, Biolegend, San Diego, CA, USA), rat anti-CD8a (100802, Biolegend, San Diego, CA, USA), rabbit anti-F4/80 (30325S, Cell Signaling, Danvers, MA, USA), rat anti-CD11b (101202, Biolegend, San Diego, CA, USA), rabbit anti-CD11c (97585S, Cell Signaling, Danvers, MA, USA), rat anti-MHCII (14-5321-85, eBioscience, Thermo Fisher Scientific, Waltham, MA, USA), rabbit anti-FoxP3 (12653, Cell Signaling, Danvers, MA, USA), rabbit anti-CD45 (ab10558, abcam, Cambridge, UK), rat anti-Ly6C (ab15627, abcam, Cambridge, UK), rat anti-Gr1 (ab 25377, abcam, Cambridge, UK), goat anti-Reelin (AF3820, R&D Systems, Minneapolis, MN, USA), goat anti-Periostin (AF2955, R&D Systems, Minneapolis, MN, USA), rabbit anti-TGFBI (ab170874, abcam, Cambridge, UK). Secondary antibodies: donkey Alexa-Fluor 488, Alexa-Fluor 647, and Cy3-conjugated secondary antibodies were purchased from Dianova (Hamburg, Germany).
Ab15627
Manufactured by Abcam
Sourced in United Kingdom, United States
Ab15627 is a recombinant protein product from Abcam. It is suitable for use in various laboratory applications, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Ab32362, by Abcam (1 mentions)
Ab10558, by Abcam (1 mentions)
Ab25377, by Abcam (1 mentions)
Ab170874, by Abcam (1 mentions)
Dm3614p, by Dianova (1 mentions)
Af1460, by R&D Systems (1 mentions)
Af3837, by R&D Systems (1 mentions)
Rat anti cd8a, by BioLegend (1 mentions)
Af3820, by R&D Systems (1 mentions)
Af2955, by R&D Systems (1 mentions)
Alexa fluor 647, by Dianova (1 mentions)
A21206, by Thermo Fisher Scientific (1 mentions)
A21434, by Thermo Fisher Scientific (1 mentions)
Mab2784, by R&D Systems (1 mentions)
Vectashield mounting medium with dapi, by Vector Laboratories (1 mentions)
A11059, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 labeled goat anti rat igg, by Thermo Fisher Scientific (1 mentions)
Anti f4 80 mca497ga, by Bio-Rad (1 mentions)
E0466, by Agilent Technologies (1 mentions)
P0397, by Agilent Technologies (1 mentions)
5 protocols using ab15627
1
Multiparametric Immunofluorescence Staining
2
Immunohistochemical Analysis of CETP in Mouse Liver
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Paraffin‐embedded sections of mouse liver (5 μm) were stained for F4/80 and human CETP (ab51771; 1/1000, Abcam) as described previously,10 Clec4f (MAB2784; 1/1000, R&D Systems), Vsig4 (AF4674; 1/1000, R&D Systems) and Ly6C (ab15627; 1/400, Abcam). For immunofluorescence staining, the secondary antibodies donkey anti‐rabbit Alexa488 (A21206; Invitrogen) and goat anti‐rat Alexa555 (A21434; Invitrogen) were used. Finally, tissue sections were mounted with VECTASHIELD® Mounting Medium with DAPI (Vector Laboratories). Positive cells were counted using a LeicaCTR5500 fluorescence microscope (Leica Microsystems GmbH). Representative pictures of immunostaining for CETP in liver sections of non‐CETP transgenic mice (APOE*3‐Leiden mice), APOE*3‐Leiden.CETP transgenic mice, and a healthy human donor are shown in Figure S1 .
3
Immunohistochemical Analysis of Liver Markers
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Liver sections were incubated with primary antibodies, followed by incubation with biotinylated secondary antibodies. The following primary antibodies were used: anti-STK25 (YSK1; sc-6865; Santa Cruz Biotechnology, Santa Cruz, CA), anti-PCNA (MA5-1158; Invitrogen), anti-Ki67 (14-5698-82; Invitrogen), anti-Gr1 (Ly6C) (ab15627; Abcam, Cambridge, UK), anti-F4/80 (MCA497GA; Bio-Rad, Hercules, CA), anti–collagen I (SAB4500362; Sigma-Aldrich), anti–α-smooth muscle actin (ab5694; Abcam), anti–4-HNE (sc-130083; Santa Cruz Biotechnology), anti-PEX5 (PA5-58716; Invitrogen), and anti-ubiquitin (ab411; Abcam). For immunohistochemical detection, anti-goat IgG (E0466; Dako, Glostrup, Denmark) and anti-mouse IgG (E0464; Dako) secondary antibodies were used, followed by horseradish-peroxidase–conjugated streptavidin (P0397; Dako) and diaminobenzidine staining (K3467; Dako). For immunofluorescence detection, Alexa Fluor-594–labeled goat anti-rat IgG (A11007; Invitrogen), Alexa Fluor-488–labeled rabbit anti-mouse IgG (A11059; Invitrogen), Alexa Fluor-594–labeled donkey anti-goat IgG (A11058; Invitrogen), and Alexa Fluor-594–labeled donkey anti-rabbit IgG (A21207; Invitrogen) secondary antibodies were used. The stained area was quantified in 5 randomly selected microscopic fields (×200) per mouse using ImageJ software.
4
Immunophenotyping of Liver Cells
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Frozen liver sections were washed and fixed with ice cold methanol for 10 min. Next, the cells were washed, permeabilized (0.1% Tween-20 in PBS, 30 min), blocked for 1 h with 1% BSA-PBS and incubated at 4 °C with mouse monoclonal rabbit anti-CHI3L1 (ab180569, 1:100, Abcam), mouse anti-F4/80 (Cat 123,106, 1:100, BioLegend) and anti-Ly6C (ab15627, 1:100, Abcam), and goat anti-CLEC4F (AF2784, 1:100, R&D Systems) antibodies diluted in Dako Antibody Diluent. After overnight incubation, cells were washed and treated with Alexa Fluor 488 goat anti-rabbit (1:1000; Invitrogen), Alexa Fluor 594 goat anti-mouse (1:1000), Alexa Fluor 488 donkey anti-goat (1:1000) or Alexa Fluor 594 donkey anti rabbit (1:1000) for 1 h at RT in the dark, followed by washing and 5 min of nuclei staining with DAPI diluted at 1:1000 in PBS.
5
Immunohistochemical Staining of Mouse Tissues
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Mouse tissues were fixed in 10% formalin and embedded in paraffin before prepared into 4-µm sections. Upon deparaffinization in xylene and rehydration through a series of diluted alcohol, tissue sections underwent antigen retrieval in boiled 10 mM citrate buffer (pH 6.0) for 10 min. After blocking endogenous peroxidase activity with 0.3% H2O2 in PBS for 10 min and non-specific binding with 5% normal goat serum for 1 h at room temperature, the tissue sections were incubated with either anti-F4/80 (ab111101, Abcam, Cambridge, MA, USA) or anti-Ly6c (ab15627, Abcam) antibody at 4 °C overnight. Following the incubation with biotinylated secondary antibody at room temperature for 30 min, the target signal was amplified using Vectastain ABC-HRP solution (Vector Labs, Burlingame, CA, USA) according to the manufacturer’s instructions and detected using Diaminobenzidine (DAB) substrate (Vector Labs).
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