Ab6789 ab6721

Manufactured by Abcam

Sourced in United Kingdom

Ab6789/ab6721 is a primary antibody for use in immunoassays. It is a polyclonal antibody raised in rabbit. The product is supplied as a liquid.

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Ab6789 Ab6721

3 protocols using ab6789 ab6721

Evaluation of Anti-Inflammatory and Antioxidant Effects

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Proteinase K and PBS tablets were ordered from MP-Biomedicals, USA. N-(1-Naphthyl) ethylenediamine dihydrochloride, trichloroacetic acid (TCA), 5,5′-dithiobis-(2-nitro benzoic acid) (DTNB), 1-chlor-2,4-dinitrobenzene (CDNB) and reduced glutathione (GSH) were obtained from Sigma Aldrich, USA. Mouse monoclonal anti-TNF-α (SC-52B83722), The 3,3′-Diaminobenzidine (DAB, SC-216567) and Avidin-biotin complex (ABC kit, SC-516216) were acquired from Santa-Cruz Biotechnology, USA. Mounting media (ab-10431) and horseradish peroxidase-conjugated secondary antibodies (ab-6789, ab-6721) were acquired from Abcam, UK. Flumazenil (FLZ) and diazepam (DZM) was procured from Shifa International Hospital, Islamabad, Pakistan. Phenyltetrazole (PTZ) was obtained from Sigma Aldrich, USA. TNF-α enzyme linked immunosorbent assay (ELISA) kit (Cat # SU-B3098) was bought from Shanghai-Yuchun Biotechnology, China. The compound 1, 3, 4-oxadiazole derivative (B3) was obtained as gift from pharmaceutical chemistry post graduate lab, Riphah International University, Islamabad, Pakistan.

Faheem M., Althobaiti Y.S., Khan A.W., Ullah A., Ali S.H, & Ilyas U. (2021). Investigation of 1, 3, 4 Oxadiazole Derivative in PTZ-Induced Neurodegeneration: A Simulation and Molecular Approach. Journal of Inflammation Research, 14, 5659-5679.

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Osteogenic Protein Expression Analysis

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The hBMMSC lysates were extracted using radioimmunoprecipitation assay buffer (Thermo Fisher Scientific, Waltham, USA), and protein quantification was performed using a BCA protein estimation kit (Thermo Fisher Scientific). Subsequently, the proteins were resolved by 10% SDS-PAGE and transferred to PVDF membranes (Thermo Fisher Scientific). The membranes were blocked with BSA (5%; Sigma) dissolved in PBS containing 0.02% Tween 20 for 60 min. The membrane was initially incubated with primary antibodies at room temperature for 1 h, followed by incubation with secondary antibodies at 4°C overnight. The antibodies used are listed as follows: anti-OCN (ab93876, 1:2000; Abcam, Cambridge, UK), anti-OPN (ab8448 1:2000; Abcam), anti-RUNX2 (ab192256, 1:500, Abcam), anti-ALP (ab67228, 1:2000; Abcam), anti-BSP (ab52128, 1:2000; Abcam), anti-Bcl-2 (ab59348, 1:5000; Abcam), anti-p-ERK1/2 (ab214362, 1:500; Abcam), anti-ERK1/2 (ab54230, 1:1000; Abcam), anti-β-actin (ab8227, 1:5000; Abcam), and HRP-conjugated goat anti-mouse/rabbit (ab6789/ab6721, 1:5000; Abcam). Protein bands were detected using Pierce™ ECL Western Blotting Substrate (Thermo Fisher Scientific), and gray images and values were recorded using a Blot Scanner (LiCor, Lincoln, USA).

Zhang Y., Dong Y., Wei Q., Zhuang Z., Liu Y., Yuan Q., He W., Jing Z., Li J., Li P., Zhang L., Hong Z., Zhang N., Wang H, & Li W. (2023). miR-126 mitigates the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by targeting the ERK1/2 and Bcl-2 pathways: Role of miR-126 in osteogenic differentiation. Acta Biochimica et Biophysica Sinica, 55(3), 449-459.

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Quantifying Viral Protein Expression

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The cell lysates from collected samples were separated on an SDS-polyacrylamide gel, and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore). The expression of oISG15 and mut-oISG15 were detected by mouse anti-HA tag antibody (ProteinTech, 51064-2-AP, 1:4,000). The viral proteins were detected by rabbit anti-VP6 (dilution = 1:1,000) and rabbit anti-NS1 antibody (dilution = 1:1,000), and both antibodies were prepared in our lab.
The HEK-293T cells were co-transfected with viral recombinant plasmids expressing VP3, VP4, VP5, NS1 protein, and oISG15-HA plasmid, respectively. Viral proteins were detected by mouse anti-Myc tag antibody (Cell Signaling Technology, 20229, 1:1,000), the ubiquitin conjugation was tested using anti-Flag antibody (ProteinTech, 66008-4-Ig, 1:5,000), and the oISG15 was detected by mouse anti-HA tag antibody (ProteinTech, 51064-2-AP, 1:5,000).
All the target proteins were visualized using a ChemiDoc MP imaging system (Bio-Rad) after incubation with the corresponding secondary antibodies (Abcam, ab6789/ab6721, 1:10,000). The band intensities of the viral proteins were measured using ImageJ software and normalized to that of β-actin.

Kang D., Zhang G., Zhang Z., Tian Z., Gao S., Liu G., Guan G., Luo J., Yin H, & Du J. (2023). Interferon-stimulated gene 15 facilitates BTV replication through interacting with the NS1 protein. Frontiers in Microbiology, 14, 1212242.

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