Retinas were processed as previously described [42 (link)] using the following antibodies: primary monoclonal antibody for CD45 (R&D Systems; MAB114, RRID:AB_357485) with secondary Alexa Fluor 594 (A11007, Invitrogen, USA); primary ionised calcium binding adaptor molecule 1 ( Iba-1; Fujifilm Cellular Dynamics; USA; 019–19741, RRID:AB_839504) with secondary Alexa Fluor 488 (A27023, Invitrogen USA) and NeuN (Millipore Sigma, USA; clone A60, Alexa Fluor 488 conjugated MAB377X) at 1:50; 1:500 and 1:100 dilutions, respectively. The antibodies were diluted in PBS with 1% normal goat serum (G9023 Sigma Aldrich, USA). Dilution for the secondary antibodies was 1:1000. The images were analysed in blinded manner.
Mab114
Manufactured by R&D Systems
Sourced in United States, United Kingdom
MAB114 is an antibody product manufactured by R&D Systems. It is a monoclonal antibody that detects an unspecified target. The antibody is supplied in liquid form.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 510 meta confocal microscope, by Zeiss (2 mentions)
A11007, by Thermo Fisher Scientific (1 mentions)
G9023, by Merck Group (1 mentions)
Iba 1, by Fujifilm (1 mentions)
Ab62623, by Abcam (1 mentions)
Ab109367, by Abcam (1 mentions)
3 3 diaminobenzidine substrate, by Vector Laboratories (1 mentions)
Igg isotype control antibody, by Santa Cruz Biotechnology (1 mentions)
Alexa 488 and 594, by Thermo Fisher Scientific (1 mentions)
Anti α sma a5228, by Merck Group (1 mentions)
Mab1398z, by Merck Group (1 mentions)
Bx53 microscope, by Olympus (1 mentions)
Ab5694, by Abcam (1 mentions)
Ab46545, by Abcam (1 mentions)
2 protocols using mab114
1
Retinal Immunofluorescence Staining Protocol
2
Histological Analysis of Tissue Samples
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For histological examination, hematoxylin and eosin (H&E) staining, and Movat pentachrome staining were performed. Anti-CD31 (MAB1398Z; Millipore, Burlington, MA), anti-alpha-smooth muscle actin (anti-α-SMA; A5228; Sigma-Aldrich, St. Louis, MO; and ab5694; Abcam, Cambridge, United Kingdom), anti-CD45 (Mab114; R&D Systems, Minneapolis, MN), anti-PRDX2 (ab109367; Abcam), anti-4-hydroxynonenal (anti-4-HNE; ab46545; Abcam), anti-DNA/RNA damage antibody (anti-8-OHG) (ab62623; Abcam), and anti-mouse macrophage/monocyte antibody (anti-MOMA2) (MCA519G; Bio-Rad, Hercules, CA) were used as primary antibodies for immunostaining. After incubation with the primary antibodies, Alexa 488 and 594 (Invitrogen, Carlsbad, CA) or biotinylated secondary antibodies with 3,3′-diaminobenzidine substrate (Vector Laboratories, Burlingame, CA) were used to visualize the antigens. Furthermore, 4′,6-diamidino-2-phenylindole (DAPI) or hematoxylin was used to label the nuclei. The negative control tissues were prepared in a similar manner using IgG isotype control antibodies (Santa Cruz Biotechnology, Dallas, TX). The immunofluorescence was imaged with an LSM 510 meta confocal microscope (Carl Zeiss, Oberkochen, Germany) or a BX53 microscope (Olympus, Tokyo, Japan).
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