Ab63917

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab63917 is a primary antibody used for the detection of a specific target protein in various laboratory applications. The core function of this product is to provide a reliable tool for researchers to identify and study the target of interest. No further interpretation or extrapolation on the intended use is provided.

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Lab products found in correlation

Image lab software, by Bio-Rad (1 mentions) Immun blot pvdf, by Bio-Rad (1 mentions) Chemidoc touch imager, by Bio-Rad (1 mentions) Mini protean tgx stain free gel, by Bio-Rad (1 mentions) Mouse anti human cd 63 clone h5c6, by BD (1 mentions) Hrp conjugated goat anti rabbit igg, by Santa Cruz Biotechnology (1 mentions) Clarity western ecl substrate, by Bio-Rad (1 mentions) Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Mouse anti human cd81, by Santa Cruz Biotechnology (1 mentions) Immobilon p membrane, by Merck Group (1 mentions) Enhanced chemiluminescence kit, by Merck Group (1 mentions) F1804, by Merck Group (1 mentions) Mouse anti β actin, by Santa Cruz Biotechnology (1 mentions) Ab31850, by Abcam (1 mentions) W4021, by Promega (1 mentions) W4011, by Promega (1 mentions) Polybrene, by Merck Group (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Histopaque 1077, by Merck Group (1 mentions) β actin hrp, by Merck Group (1 mentions)

4 protocols using ab63917

Western Blot Analysis of Extracellular Vesicles

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Vesicles recovered from conditioned medium from 20 × 10⁶ cells (2 K, 10 K and 100 K pellets), equivalent volumes of cell-free medium (2 K, 10 K and 100 K pellets) or 15 μl of the iodixanol gradient fractions were loaded on 4–15% or 10% Mini‐Protean® TGX Stain‐Free™ gels (Bio‐Rad), under non‐reducing conditions. Transferred membranes (Immun‐Blot PVDF Bio‐Rad) were developed (e.g. using Clarity western ECL substrate (Bio-Rad) or SuperSignal West Pico Plus Chemiluminescent Substrate (Thermo Fisher) and a ChemiDoc Touch imager (Bio‐Rad) or by standard film exposure). Intensity of the bands was quantified using ImageLab Software (Bio‐Rad). Antibodies for WB were mouse anti‐human CD63 (clone H5C6, BD Bioscience #557305), mouse anti-human CD81 (Santa Cruz sc-23692), goat anti-AChE (ab31276, Abcam) and anti-HIV-1 p24 Monoclonal (183-H12-5C, NIH AIDS reagent program) or Anti-HIV1 p55 + p24 + p17 (ab63917, Abcam). Secondary antibodies included HRP-conjugated goat anti-rabbit IgG (Santa Cruz, sc-2004) and HRP-conjugated m-IgG-k BP (Santa Cruz, sc-516102) (JHU), or (CT lab) HRP-conjugated anti-mouse (Jackson Immunoresearch, 115–035-146) and anti-goat (Jackson, 705–035-147).

Liao Z., Jaular L.M., Soueidi E., Jouve M., Muth D.C., Schøyen T.H., Seale T., Haughey N.J., Ostrowski M., Théry C, & Witwer K.W. (2019). Acetylcholinesterase is not a generic marker of extracellular vesicles. Journal of Extracellular Vesicles, 8(1), 1628592.

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Quantitative Western Blot Analysis

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Whole-cell extracts were prepared by lysing cells, boiling the equivalent protein content in sodium dodecyl sulfate sample buffer, resolution by SDS/PAGE, and transfer to an Immobilon-P membrane (Millipore, Germany). The membranes were incubated with primary antibodies, mouse anti-β-actin (sc-47778, Santa Cruz Biotechnology, Inc. USA, 1:5000), mouse anti-p24 (ab63917, Abcam, USA, 1:2500), rabbit anti-Spasitn (ab31850, Abcam, USA, 1:1000), rabbit anti-CD4 (19068-1-AP, Proteintech Group, Inc. USA, 1:1000), rabbit anti-CHMP1B (14639-1-AP, Proteintech Group, Inc. USA, 1:2000), and rabbit anti-IST1 (51002-1-AP, Proteintech Group, Inc. USA, 1:5000), mouse anti-FLAG (F1804, Sigma-Aldrich, USA, 1:5000) followed by the horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (W4011, Promega, USA, 1:10000) or anti-Mouse IgG secondary antibodies (W4021, Promega, USA, 1:10000). Immunoreactive proteins were detected using an enhanced chemiluminescence kit (Millipore, Germany). The intensities of Western blot bands were quantified using digital images and Image J software after subtraction of the background.

Shen W., Liu C., Hu Y., Ding Q., Feng J., Liu Z, & Kong X. (2023). Spastin is required for human immunodeficiency virus-1 efficient replication through cooperation with the endosomal sorting complex required for transport (ESCRT) protein. Virologica Sinica, 38(3), 448-458.

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Antibody-based detection of HIV proteins

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The following antibodies have been used in the study: MATR3 (A300-590A, Bethyl Laboratories, 1:2,000 dilution for IB), HIV-1 p55 and p24 (HIV-1 p24 [1941], sc-65462, Santa Cruz, or anti-HIV-1 p55⁺ p24⁺ p17 antibody ab63917, Abcam, 1:200). Hsp90 (catalogue number ALX-804-808, Enzo Life Sciences), PARP (catalogue number ALX-210-302, Enzo Life Sciences), β-actin HRP (Sigma, AC-15, 1:10,000), PSF (B92 Sigma P2860, 1:1,000), cyclin T1 (C-20-SC-8128, 1:200), polypyrimidine tract binding protein PTB (rabbit polyclonal produced in-house, 1:1,000), flag tag (Sigma, 1:5,000), and vimentin (Cell Signaling, 1:1,000).
Other reagents included Histopaque-1077 (10771, Sigma), Polybrene (Sigma-H9268), phytohemagglutinin (PHA L1668-5MG, Sigma), interleukin-2 (H7041, Sigma), TNF-α (T0157, Sigma), puromycin (ant-pr-1, InvivoGen), blasticidin (ant-bl-1, InvivoGen), SAHA (SML0061, Sigma), disulfiram (D2950000, Sigma), romidepsin (S3020, Selleckchem), and JQ1 (2091-1, BioVision). IngenolB was kindly donated by Luiz F. Pianowski, Kyolab/Amazônia Fitomedica-mentos, Valinhos, Sao Paulo, Brazil. Human CD3 (IMI1304) and CD28 (IMI1376) antibodies were obtained from Analis (Belgium).

Sarracino A., Gharu L., Kula A., Pasternak A.O., Avettand-Fenoel V., Rouzioux C., Bardina M., De Wit S., Benkirane M., Berkhout B., Van Lint C, & Marcello A. (2018). Posttranscriptional Regulation of HIV-1 Gene Expression during Replication and Reactivation from Latency by Nuclear Matrix Protein MATR3. mBio, 9(6), e02158-18.

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Western Blot Detection of Fluorescent Proteins

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Rabbit monoclonal antibody against GFP (EPR14104, ab183734), rabbit polyclonal antibody against mCherry (ab167453), and rabbit polyclonal to HIV-1 gag (p55 + p24 + p17, ab63917) were purchased from Abcam (UK). Horse-radish peroxidase-conjugated goat anti-rabbit was obtained from Agilent Dako (USA). NuPAGE™ 4-12% Bis-Tris Protein Gels and iBlot™ Transfer Stacks were purchased from ThermoFisher Scientific (USA). Immunoblotted proteins were detected using the Enhanced Chemiluminescence detection system (Amersham, UK) and ChemiDoc XRS+ (Bio-Rad, USA). Pan-caspase inhibitor zVAD-FMK was purchased from Enzo Life Sciences (USA) and GrM inhibitor AcKVPL-CMK from Peptanova (Germany).

Saccon E., Mikaeloff F., Figueras Ivern P., Végvári Á., Sönnerborg A., Neogi U, & van Domselaar R. (2021). Cytotoxic Lymphocytes Target HIV-1 Gag Through Granzyme M-Mediated Cleavage. Frontiers in Immunology, 12, 669347.

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