Sc 1615

Manufactured by Abcam

Sourced in United States

Sc-1615 is a primary antibody that recognizes CD4, a transmembrane glycoprotein expressed on the surface of helper T cells. It functions as a co-receptor for the T cell receptor, facilitating the activation of T cells in response to antigen presentation.

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Lab products found in correlation

4 protocols using sc 1615

Antibody Procurement and Validation

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Antibodies were purchased from Santa Cruz Biotechnology (IRF4, sc-6059; actin, sc-1615; PGC-1α, sc-13067), Abcam (UCP1, AB10983; Mito profile, AB110413; Aconitase, AB110321), and Millipore (PGC-1α, ST1202-1SET).

Kong X., Banks A., Liu T., Kazak L., Rao R.R., Cohen P., Wang X., Yu S., Lo J.C., Tseng Y.H., Cypess A.M., Xue R., Kleiner S., Kang S., Spiegelman B.M, & Rosen E.D. (2014). IRF4 is a key thermogenic transcriptional partner of PGC-1α. Cell, 158(1), 69-83.

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Western Blot Immunodetection Protocol

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Proteins were resolved by Mini Gel SDS-PAGE (Biorad^® system) and transferred to PVDF membrane (Immobilon-P^®- Millipore^®) according to standard procedures. Blocking and antibody incubations were performed in TBS - 0.2% Tween-20^® 5% milk (Marvel^®). The following primary antibodies were used: anti-Ku80, (1:1000)^{38 (link)}, anti-actin (1:5000, Santa-Cruz^® sc-1615), anti-H2AX-P (1:1000, Abcam® ab11174), anti-H3 (1:2000, Abcam^® ab12079), anti-flag (1:2000, SIGMA^® F1804 and F2425), anti-mono-ADPr AbD33205 (1:1000)^{34 (link)}, Anti-H3S10-p (1:1000, Bethyl^® A301-844A-T, and Abcam^® ab5176). Global ADP-ribosylation sigma was detected using the reagent anti-PAN-ADPr (1:2000, Merk^® MABE1016). Appropriate HRP-conjugated secondary antibodies were used anti-mouse (1;10000, DAKO^®), anti-rabbit (1;10000, DAKO^®), anti-goat (1;10000, DAKO^®) and anti-human (1:5000)^{34 (link)}. Immuno-reactive bands were detected by chemo-luminescence induced by Immobilon^® western substrate (Millipore^®), detected with the LI-COR^® Odyssey-Fc machine and quantified using Image-Studio^® software.

Brustel J., Muramoto T., Fumimoto K., Ellins J., Pears C.J, & Lakin N.D. (2022). Linking DNA repair and cell cycle progression through serine ADP-ribosylation of histones. Nature Communications, 13, 185.

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Exosome and Cell Lysate Characterization

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Cells were lysed in lysis buffer supplemented with PIs, for 15 min at 4 °C, centrifuged at 12000g for 15 min at 4 °C and the supernatant collected and stored at −20 °C until further use. EV (exosomes) and cell lysate protein concentration was measured using Pierce BCA Protein Assay Kit (Thermo Scientific MA, USA).
For immunoblotting, total EV, exosomal, and viral proteins or 30 μg of cell lysates were separated by 12% SDS-PAGE and transferred to PVDF membrane (Millipore). Goat polyclonal antibodies against Hsc70 (sc-1059, Santa Cruz Biotechnology, CA, USA), actin (sc-1615), rabbit polyclonal against calnexin (sc-11,397), CD63 (sc-15,363), alpha-tubulin (ab4074, Abcam, UK), VSV-G (ab1874, Abcam), HIV-1 Nef (NIH AIDS Reagent Program, 2949), and HIV-1 gp120 (NBP1–76371, Novus Biologicals, CO, USA); or mouse monoclonal against acetylcholinesterase (AChE, MAB303, Millipore), flotillin, (610820, BD Bioscience, NJ, USA), GFP (sc-9996), p24/Gag (ab9071, Abcam), cytochrome c (556432, BD Biosciences), and Alix (2171S, Cell Signaling, MA, USA) were used as the primary antibodies; and appropriate HRP-conjugated anti-goat, anti-rabbit, or anti-mouse (Jackson Immunoresearch, PA, USA) were used as the secondary antibodies. Membranes were developed by SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific).

Dominkuš P.P., Ferdin J., Plemenitaš A., Peterlin B.M, & Lenassi M. (2017). Nef is secreted in exosomes from Nef.GFP-expressing and HIV-1-infected human astrocytes. Journal of neurovirology, 23(5), 713-724.

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Western Blot Analysis of Cellular Markers

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For western blot analysis, the fractional homogenate samples (cytosolic and membrane bound) were prepared in the sample buffer (100 mM Tris-Cl [pH 6.8], 4% [w/v] SDS, 0.2% [w/v] bromophenol blue, 20% [v/v] glycerol, and 200 mM dithiothreitol [DTT]). Samples were then electrophoresed on 10% SDS-PAGE gels and the proteins were transferred to a nitrocellulose membrane. Immunoblotting was performed using primary antibodies against CD105/endoglin (0.25 μg/mL, R&D systems; AF1097), Aβ (1:1,000, 6E−10; BioLegend 39320), anti-ZO1 (2 μg/mL, Thermo Fisher Scientific; 61-7300), anti-GAPDH antibody (1:100, Abcam; ab181602), anti-actin (1:1000, Santa Cruz; sc1615 and anti-VEGFa (1 μg/mL, Abcam; ab46154). After the overnight incubating with primary antibodies at 4°C, the membranes were washed three times with 1× PBS solution for 15 min each and then incubated with the secondary antibodies (1:10,000) for 1 h at room temperature followed by washing three times with 1× PBS. The signal intensities on the membrane were imaged using the Odyssey infrared image system (LICOR), and relative levels of immunoreactivity were analyzed by the Image Studio Lite Software. The blots shown are representative of n = 6 per group.

Singh C.S., Johns K.M., Kari S., Munro L., Mathews A., Fenninger F., Pfeifer C.G, & Jefferies W.A. (2024). Conclusive demonstration of iatrogenic Alzheimer’s disease transmission in a model of stem cell transplantation. Stem Cell Reports, 19(4), 456-468.

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