The slides of cells were embedded in paraffin according to standard pathology protocols. After antigen retrieval and permeabilization with 0.3% Triton X‐100, tissues were incubated incubated at 4°C overnight with the recommended concentrations of primary Abs (mouse anti‐human antibodies, including C5a (ab11877, Abcam), CCL2 (MABN712, Millipore), and rabbit anti‐human antibodies, such as C5aR1 (ab59390, Abcam), CCR2(NBP1‐48337, Novusbio)) by the manufacturer. After washing, slides were incubated with selected secondary Abs (AF594‐ goat anti‐mouse IgG (ab150116, Abcam), AF488‐ goat anti‐rabbit IgG (ab150077, Abcam), AF647‐ donkey anti‐goat IgG (ab150131, Abcam)) for 40 min at room temperature in the dark, correctly matched to the appropriate species. DAPI mounting medium (Vector Laboratories) was used for cell nuclei staining. Finally, slides were viewed under an imaging fluorescence microscope (Olympus BX51; Olympus).
Mabn712
Manufactured by Merck Group
Sourced in Germany
MABN712 is a laboratory centrifuge designed to separate components of a liquid mixture based on their relative densities. It features a motor-driven rotor that spins at high speeds, creating a centrifugal force that causes the denser components to move outward, effectively separating them from the less dense components. The MABN712 is a essential piece of equipment for various scientific and industrial applications that require the separation and isolation of substances.
Automatically generated - may contain errors
Lab products found in correlation
Ab150116, by Abcam (2 mentions)
Ab150131, by Abcam (1 mentions)
Ab11877, by Abcam (1 mentions)
Ab59390, by Abcam (1 mentions)
Dapi mounting medium, by Vector Laboratories (1 mentions)
Nbp1 48337, by Novus Biologicals (1 mentions)
Ab150077, by Abcam (1 mentions)
Ab182032, by Abcam (1 mentions)
Ab32457, by Abcam (1 mentions)
Ab85034, by Abcam (1 mentions)
Ab203883, by Abcam (1 mentions)
Ab9498, by Abcam (1 mentions)
Ab104139, by Abcam (1 mentions)
Ab33471, by Abcam (1 mentions)
Toto 3, by Thermo Fisher Scientific (1 mentions)
Tcs sp2, by Leica (1 mentions)
Ab9722, by Abcam (1 mentions)
3 protocols using mabn712
1
Immunofluorescence Staining of Paraffin-Embedded Tissues
2
Kidney Tissue Immunofluorescence Staining
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Immunofluorescence staining was performed in kidney tissue embedded in paraffin according to standard pathology protocols. The primary antibodies used were mouse anti-human antibodies CD14 (ab182032, Abcam), CD31 (ab9498, Abcam), CCL2 (MABN712, Millipore), and rabbit anti-human antibodies CD16 (ab203883, Abcam), CD31 (ab32457, Abcam), CX3CL1 (ab85034, Abcam). The slides were placed in a wet chamber followed by the addition of the appropriate primary antibodies at the concentration recommended by the manufacturer (double staining) and incubated overnight at 4°C. The slides were rinsed three times with PBS, and a 1:200 dilution of Alexa Fluor®594 goat anti-mouse IgG antibody (ab150116, Abcam) and Alexa Fluor®488 goat anti-rabbit IgG antibody were applied and incubated at 37°C (30 minutes) in the dark. DAPI mounting medium (ab104139, Abcam) was used for nuclear staining. Finally, the slides were viewed under fluorescent microscopy (Olympus BX51; Olympus, Tokyo, Japan).
3
Immunofluorescence Analysis of Inflammatory Markers
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Immunofluorescence analysis was performed using cell coverslip. Briefly, the cover slip was fixed in 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton-x 100 for 10 min, and blocked with 2% bovine serum albumin (BSA) at room temperature for 1 h, and then incubated with primary antibodies monocyte chemoattractant protein (MCP-1) (MABN712, 1:200, Millipore, Germany), macrophage inflammatory protein-1α (MIP-1α) (MAB66251, 1:100, RD Biosciences, USA), TNF-α (SC1069, 1:100, Sant Crus, USA), IL-1β (ab9722, 1:200, Abcam, USA), and IL-10 (ab33471, 1:300, Abcam, USA) overnight. Appropriate secondary antibodies were used by FITC-conjugation (goat anti mouse, 1:200, Invitrogen, USA; rabbit anti rat, 1:20, DAKO, Japan; chicken anti goat, 1:200, Invitrogen; chicken anti rabbit, 1:200, Invitrogen; rabbit anti rat, 1:20, DAKO; respectively). The coverslips were counterstained with Toto3 (1:500, Invitrogen, USA) for nucleus. Slides were examined by confocal microscopy (Leica TCS SP2, Germany) and images were analyzed by F'cell software.
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