Mouse brains from NCLX-nKO (NCLXfl/fl x Camk2a-Cre) and control (Camk2a-Cre) mice were prepared for immunohistochemistry.18 (link) In brief, brains were fixed in 4% paraformaldehyde overnight, embedded in paraffin, and sectioned. 6-μm thick coronal sections were then deparaffinized, hydrated, and blocked in 2% fetal bovine serum. The sections were incubated with primary antibody overnight at 4°C. The primary antibodies and dilutions were as follows: HT7 dilution 1:150 (Thermo Fisher Scientific, Catalog # MN1000), phospho-tau (pThr231) monoclonal AT180 dilution 1:50 (Thermo Fisher Scientific, Catalog # MN1040) and anti-4 hydroxynonenal antibody (4-HNE) dilution 1:20 (Abcam, Catalog # ab48506). The sections were then incubated with secondary antibodies and developed using the Vector Elite ABC (Avidin-Biotin Complex) system (Vector Laboratories Inc., Burlingame, CA).
Anti 4 hydroxynonenal antibody 4 hne
Manufactured by Abcam
Sourced in Japan, United States
Anti-4 hydroxynonenal antibody (4-HNE) is a laboratory reagent used for the detection and quantification of 4-hydroxynonenal, a lipid peroxidation product. This antibody can be utilized in various analytical techniques, such as Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA), to study the role of 4-HNE in biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Sulforaphane, by Merck Group (1 mentions)
Anti glyceraldehyde 3 phosphate dehydrogenase gapdh, by Santa Cruz Biotechnology (1 mentions)
Anti erk, by Cell Signaling Technology (1 mentions)
Anti p p38 thr180 tyr182, by Cell Signaling Technology (1 mentions)
Anti p65, by Santa Cruz Biotechnology (1 mentions)
Anti p erk thr202 tyr204, by Cell Signaling Technology (1 mentions)
Anti sapk jnk, by Cell Signaling Technology (1 mentions)
Anti p21, by Santa Cruz Biotechnology (1 mentions)
Anti p38, by Cell Signaling Technology (1 mentions)
Anti iκbα, by Cell Signaling Technology (1 mentions)
Quercetin, by Merck Group (1 mentions)
Resveratrol, by Merck Group (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Vimentin, by Agilent Technologies (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
4 protocols using anti 4 hydroxynonenal antibody 4 hne
1
Immunohistochemical Analysis of Mouse Brains
2
Quantifying Antioxidant Enzymes and Oxidation
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SDS-PAGE electrophoresis and transfer to PVDF membrane were performed as described [33 (link)]. Rabbit anti-SOD2 (Mitochondrial matrix MnSOD, Abcam, Paris, France), mouse anti-N(epsilon)-(hexanoyl)lysine (HEL, Cosmobio, Tokyo, Japan) and anti—4 Hydroxynonenal antibody (4-HNE, Abcam, Paris, France) monoclonal antibodies were used for antioxidant enzyme and oxidation product detection, respectively. Detection of fluorescent secondary antibodies (Rabbit anti-mouse 680 nm and Goat anti-rabbit 790 nm) was performed using a LiCor Odyssey apparatus (Licor Biotechnology, Bad Homburg, Germany) with dual wavelength fluorescence detection (700 and 800 nm).
3
Oxidative Stress Pathway Analysis
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Anti-poly (ADP-ribose) polymerase (PARP), anti-active caspase-3, anti-light chain 3B (LC3B), anti-phospho-p65 (Ser536) (p-p65), anti-acetyl-p65 (Lys310), anti-Nrf2, anti-IκBα, anti-p-p38 (Thr180/Tyr182), anti-p38, anti-p-p46/54 SAPK/JNK (Thr183/Tyr185), anti-SAPK/JNK, anti-p-ERK (Thr202/Tyr204), and anti-ERK antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-Del-1, anti-p21, anti-p65, and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-8-hydroxy-2′-deoxyguanosine (8-OHdG) antibody was purchased from Bioss (Woburn, MA, USA). The anti-4-hydroxynonenal (4-HNE) antibody was purchased from Abcam (Boston, MA, USA). Lipopolysaccharide (LPS), quercetin, resveratrol, and sulforaphane were obtained from Sigma-Aldrich (St. Louis, MO, USA).
4
Histopathological and Molecular Characterization of Tumors
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The tumors excised from mice were fixed in 10 % paraformaldehyde for 48 h and paraffin-embedded. Consecutive sections of 4 microns thick were cut and processed for hematoxylin and eosin staining and immunohistochemistry for Ki-67 (used at 1:100; catalog No: M7240, Dako) and cleaved caspase-3 (used at 1:200, catalog No: 9661, Cell Signaling Technology). Anti-4 Hydroxynonenal (4-HNE) antibody (catalog No: ab46545, used at 1:100) and CD31 antibodies were from Abcam (used at 1: 200, catalog No. 28364). Vimentin (used at 1:100, catalog No: M0725) antibody was obtained from Dako and staining was performed according to the manufacturer’s instruction. The positive cells stained brown. The slides were examined under a light microscope, and representative pictures were taken from a minimum of 4–5 slides from each group [23 (link)]. The quantification of the stain intensity was performed by assigning a score of 0–1 for no or weak stain; 2 for moderate stain and 3 for strong stain. All slides were examined in a blinded manner.
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