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Sc 816

Manufactured by Cell Signaling Technology
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Sourced in United Kingdom

Sc-816 is a primary antibody that binds to and detects Ikaros, a transcription factor involved in lymphocyte development and differentiation. It can be used for Western blotting and immunoprecipitation applications.

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Lab products found in correlation

Ab53280, by Abcam (1 mentions) Ab52635, by Santa Cruz Biotechnology (1 mentions) Trans blot turbo transfer system, by Bio-Rad (1 mentions) Clarity western ecl, by Bio-Rad (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Alexa 594, by Thermo Fisher Scientific (1 mentions) Envision system, by Agilent Technologies (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

AR (M-20) sc-816 (2 citations)

4 protocols using sc 816

1

Xenograft Tumor Growth Inhibition

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LNCaP-abl cells (1.5 × 107) were mixed with an equal volume of Matrigel (50 μl) and injected subcutaneously into the flank of 6-week old NOD scid gamma (NSG) male mice. When tumors reached an average of 100 mm3, mice were randomized into two groups of five and treated by intraperitoneal injection of DMSO or 50 mg/kg MPC6 twice a week. Tumor volume was measured twice weekly. After 3.5 weeks of treatment, mice were weighed and sacrificed. Xenograft studies were performed at the Memorial Sloan-Kettering Cancer Center, and was approved by their Institutional Animal Care and Use Committee. For immunohistochemistry, xenograft tumors were extracted and fixed in 4% (vol/vol) paraformaldehyde, embedded in paraffin, and sectioned. Tissue sections were prepared by NYUMC histopathology core and stained as previously described (19 (link)) using antibodies against AR (N-20, Santa Cruz Biotechnology; sc-816), and Ki67 (Cell Signaling; cat # 9027).
Wang Y., Dehigaspitiya D.C., Levine P.M., Profit A.A., Haugbro M., Imberg-Kazdan K., Logan S.K., Kirshenbaum K, & Garabedian M.J. (2016). Multivalent peptoid conjugates which overcome enzalutamide resistance in prostate cancer cells. Cancer research, 76(17), 5124-5132.
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2

Protein Isolation and Western Blot Analysis

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Organoids were collected and protein was isolated using RIPA-buffer (#sc-24948) according to the manufacturer’s protocol. Protein concentration was measured using the Pierce BCA Protein Assay Kit (#23225). Proteins were blotted on nitrocellulose membranes using a Trans-Blot TurboTM Transfer System (Bio-Rad, Hercules, CA, USA). Blots were washed with 1×TBS and blocked with 5% milk for 1 h. Membranes were incubated with primary antibodies for CK8 (Abcam, Cambridge, UK, ab53280, 1:10,000), CK5 (Abcam, Cambridge, UK, ab52635, 1:10,000), AR (Santa Cruz, sc-816, 1:200), GAPDH (Cell Signaling, Danvers, MA, USA, 5174, 1:1000), or Tubulin (Sigma, T5168, 1:1000) O/N. Membranes were washed and incubated with secondary antibodies (Cell Signaling, 7076S & 7074S, 1:1000) for 1 h. Protein was detected using Bio-Rad Clarity Western ECL (Bio-Rad, Hercules, CA, USA), according to manufacturer’s protocol.
Marhold M., Udovica S., Topakian T., Horak P., Horvat R., Tomasich E., Heller G, & Krainer M. (2022). MALAT1 Fusions and Basal Cells Contribute to Primary Resistance against Androgen Receptor Inhibition in TRAMP Mice. Cancers, 14(3), 749.
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3

Immunohistochemical and Immunofluorescent Analysis

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Formalin-fixed/paraffin-embedded specimens were sectioned at 5-μm thickness and mounted on positively charged slides. Sections were stained with hematoxylin and eosin (H&E) for histology analysis. Immunohistochemistry was performed using a standard protocol as previously described [28] (link). For detection of AR and Src expression, primary antibodies for AR (Santa Cruz Biotechnology, SC-816, 1:200) and Src (Cell Signaling, 2109, 1:250) were used and detected with the EnVision+ system (Dako). For immunofluorescent analysis, sections were incubated with primary antibodies against vimentin (Novus Biologicals, NB300-223, 1:250), E-cadherin (Cell Signaling, 3195, 1:250), CK5 (BioLegend, 905501, 1:500), or CK8 (BioLegend, 904801, 1:1000). Expression was detected by Alexa-594– or Alexa-488–conjugated secondary antibodies (Molecular Probes; 1:1000) and DAPI (Vector Laboratories) nucleus staining. The images were taken by a fluorescent microscopy with a CCD camera.
Li Q., Ingram L., Kim S., Beharry Z., Cooper J.A, & Cai H. (2018). Paracrine Fibroblast Growth Factor Initiates Oncogenic Synergy with Epithelial FGFR/Src Transformation in Prostate Tumor Progression. Neoplasia (New York, N.Y.), 20(3), 233-243.
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4

Immunofluorescent Staining of Cell Cultures

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Cells were plated at 60,00 cells per well in a 2 chambered glass slide. The following day, the chambers were removed and the slides were rinsed with pre-warmed 1x PBS, followed by 4% paraformaldehyde fixation, washed in PBS + .3% Triton-X100 (PBS-TX) and incubated in PBS-TX + 1% BSA for 1 h at room temperature. Primary antibodies (AR N-20, Santa Cruz Biotechnology #sc-816 and Tubulin, Cell Signaling #3873) were diluted in PBS-TX +1% BSA overnight at 4C. Slides were washed in PBS-TX, followed by the secondary antibody (Alexa Fluor 488 #A11008 and Alexa Fluor 647 #A21235, Invitrogen) for 1 h at room temperature (in the dark). Vectashield with DAPI was applied and allowed to dry before visualization using an EVOS FL2 Auto Imaging System.
Katleba K., Lombard A.P., Tsamouri M.M., Baek H.B., Nishida K.S., Libertini S.J., Platero A.J., Ma A.H., Pan C.X., Ghosh P.M, & Mudryj M. (2021). Depletion of androgen receptor low molecular weight isoform reduces bladder tumor cell viability and induces apoptosis. Cancer letters, 504, 49-57.
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