- PA5-22990 - Invitrogen Antibodies MAP1LC3A antibody

Zhang S, He W, Li A, Zhao C, Chen Y, Xu C, Zhang Q, Zheng D, Chen M, Miao H, Huang Y
Molecular medicine (Cambridge, Mass.) 2022 May 3;28(1):46

Singh NP, Miranda K, Singh UP, Nagarkatti P, Nagarkatti M
Toxicology 2018 Dec 1;410:49-58

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of LC3A using a polyclonal antibody (Product # PA5-22990).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of LC3A in HeLa and Neuro2A cells. Samples were incubated in LC3A polyclonal antibody (Product # PA5-22990) using a dilution of 2 µg/mL followed by an anti-rabbit HRP secondary antibody. Cells treated with or without 50 µM chloroquine for 24 hours was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 µm PVDF membrane and blocked in 5% non-fat milk in TBST. Detection: chemiluminescence. Note the detection LC3 II upon chloroquine treatment.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of LC3A in Human brain lysate. Sample was incubated in LC3A polyclonal antibody (Product # PA5-22990).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of LC3A in 0.5 mg/mL Neuro2A lysate. Samples were incubated in LC3A polyclonal antibody (Product # PA5-22990). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-LC3A Rabbit Polyclonal Antibody (Product # PA5-22990) and a 15 kDa band corresponding to LC3A was observed across cell lines tested and increased upon Chloroquine treatment. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with Chloroquine (50uM for 12 Hours) (Lane 2), HCT 116 (Lane 3) and HCT 116 treated with Chloroquine (50uM for 12 Hours) (Lane 4) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of LC3A in HeLa cells. Samples were incubated in LC3A polyclonal antibody (Product # PA5-22990) followed by DyLight 488 (green). Cells were treated overnight with 50 µM chloroquine to induce autophagosome formation. Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of LC3A in formalin-fixed paraffin-embedded tissue section of mouse brain. Samples were incubated in LC3A polyclonal antibody (Product # PA5-22990) using a dilution of 1:300 followed by a HRP-labeled secondary antibody and DAB reagent. The sections/nuclei were further counterstained with hematoxylin. Note the diffused cytoplasmic staining of LC3 in all of the cells with highest positivity in various neurons.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of LC3A in formalin-fixed paraffin-embedded tissue section of mouse liver. Samples were incubated in LC3A polyclonal antibody (Product # PA5-22990) using a dilution of 1:300 followed by a HRP-labeled secondary antibody and DAB reagent. The sections/nuclei were further counterstained with hematoxylin. Note the diffused cytoplasmic staining of LC3 in all of the hepatocytes and other liver cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of LC3A using a polyclonal antibody (Product # PA5-22990).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Risperidone causes an inhibition of autophagy and a promotion of apoptosis in femur tissues of mice. Mice were treated with 0, 0.3, 0.45, 0.6, 0.75 mg/kg of Risperidone. A representative micro-CT 3D and plan scan images of cross-sectional area of femur tissues. B statistical analysis of BMD in femur tissues of mice assessed by micro-CT. C statistical analysis of BMC in femur tissues of mice. D representative images of HE staining in femur tissues of mice. E Western blot analysis of autophagy-related proteins (LC3 II/I, Beclin1, and p62) expression in femur tissues of mice. The band intensity was assessed. F Western blot analysis of apoptosis-related proteins (cleaved PARP1, cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9). Different numbers represent different concentrations of Risperidone (mg/kg). G co-localization of LC3 II (red), LC3 I (red), Beclin1 (red), and p62 (red) with the osteoblast marker RUNX2 (green) by double-labeled immunofluorescence, wherein the nucleus is labeled by DAPI (blue). * p < 0.05, compared with the treatment of 0 mg/kg of Risperidone. The experiments are repeated 3 times

PA5-22990