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Sc 8033

Manufactured by Santa Cruz Biotechnology
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Sourced in United States

SC-8033 is a laboratory instrument designed for centrifugation applications. It is a high-speed, temperature-controlled centrifuge capable of separating and isolating various biological samples, such as cells, proteins, and nucleic acids. The product specifications and technical details are available upon request.

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Lab products found in correlation

Sc 514592, by Santa Cruz Biotechnology (2 mentions) Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Prolong gold mounting medium, by Thermo Fisher Scientific (1 mentions) Sp5 2 confocal microscope, by Leica (1 mentions) Fsx100, by Olympus (1 mentions) Sc 8038, by Santa Cruz Biotechnology (1 mentions) Glucagon, by Santa Cruz Biotechnology (1 mentions) Alexa flour conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Insulin, by Santa Cruz Biotechnology (1 mentions) Vimentin, by Cell Signaling Technology (1 mentions)

7 protocols using sc 8033

1

Immunofluorescent Imaging of Pancreatic Islets

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Tissue samples of pancreatic islets were embedded in optimal cutting temperature compound and snap-frozen in liquid nitrogen. Tissue sections (4 μm) were fixed in ethanol–acetic acid fixative solution for 2–10 min and stained with anti-insulin antibody (1:100, SC-8033, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) with secondary antibody (1:400, Fab’2 Donkey anti-Mouse IgG-AF594, Jackson ImmunoResearch Inc. West Grove, PA, USA), DAPI (Invitrogen, Waltham, MA, USA), and Alexa Fluor 488 Phalloidin (Invitrogen, Waltham, MA, USA) overnight at 4 °C in a humidified chamber. After three washes, slides were stained with DAPI and mounted with ProLong Gold mounting medium (Invitrogen, Waltham, MA, USA). Confocal imaging was performed using a Leica SP5 II confocal microscope.
Li T.T., Lin C.L., Chiang M., He J.T., Hung C.H, & Hsieh C.C. (2023). Cytokine-Induced Myeloid-Derived Suppressor Cells Demonstrate Their Immunoregulatory Functions to Prolong the Survival of Diabetic Mice. Cells, 12(11), 1507.
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2

Histological Assessment of Insulitis in Pancreatic Tissues

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Pancreatic tissues were fixed in 10% formaldehyde, 5-μm paraffin sections were made, and stained with hematoxylin and eosin (H&E) to assess insulitis. Stained sections were analyzed using a 0-4 insulitis severity grading system as described in our earlier studies[45 (link),49 (link),50 ]. In some experiments, paraffin sections of pancreatic tissues were stained using anti-insulin (mouse monoclonal, sc-8033 Santa Cruz Biotechnology) and/or anti-glucagon (rabbit polyclonal; Santa Cruz Biotechnology antibodies, followed by Alexa fluor 488- and 568- linked secondary antibodies and DAPI, and scored for insulitis based on DAPI-positive cells in islet areas and insulin positivity. In some experiments, insulin positive islets among total islet structures in different groups were compared. In some experiments, control (GFP expressing)-MSCs were injected into pre-diabetic and early hyperglycemic mice as described above and euthanized after 48h along with non-injected control mice, pancreatic tissues were subjected to cryo-sectioning. Intermittent sections were stained using aforementioned anti-insulin antibody followed by Alexa-flour 568 linked anti-mouse IgG secondary antibody, or with the secondary antibody alone, and DAPI. Stained sectioned were examined for the presence of GFP+ cells in islet area with and without immune cell infiltration based on insulin and DAPI staining.
Gaudreau M.C., Gudi R.R., Li G., Johnson B.M, & Vasu C. (2021). Gastrin producing syngeneic mesenchymal stem cells protect non-obese diabetic mice from type 1 diabetes. Autoimmunity, 55(2), 95-108.
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3

Quantifying Insulin and IR Expression

Cited 1 time
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Expression of insulin and IRs measurement in the adipose and muscle tissues was performed using the IHC staining method. The adipose and skeletal muscle tissues embedded with paraffin were cut into 5-mm thickness. The slides were incubated in an oven at 40°C overnight. Then, they were then stained with an anti-insulin primary antibody (SC-8033, Santa Cruz Biotechnology, USA) or anti-IRs-1 primary antibody (SC-8038, Santa Cruz Biotechnology) in 2% bovine serum albumin (BSA) (1:500) [15 ]. In addition, they were stained with secondary antibody goat anti-mouse IgG fluorescein isothiocyanate (02-18-06, KPL, USA) or goat anti-mouse IgG tetramethylrhodamine isothiocyanate (ab6768, Abcam, UK) in 2% of BSA (1:1500) [9 (link)]. They were washed with phosphate-buffered saline (PBS) (pH 7.4) 3 times, dried, and then observed under a fluorescence microscope (FSX 100, Olympus, Japan). The expression of insulin or IRs was analyzed and presented as intensity/mm2 (Int/mm2). The intensity for each slide was measured using the Fiji-ImageJ2 software (https://imagej.net/software/fiji/).
Retnaningtyas E., Susatia B., Arifah S.N, & Rahayu Lestari S. (2022). The improvement of insulin level after hydrogen-rich water therapy in streptozotocin-induced diabetic rats. Veterinary World, 15(1), 182-187.
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4

Immunohistochemical Analysis of Pancreatic Markers

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The heat-induced epitope retrieval using microwave in citrate buffer (10 mM citric acid, 0.05% Tween 20, pH = 6.0, Merck, Darmstadt, Germany) was performed. When needed, tissue sections were incubated in 3% H2O2 in TBST (Tris-buffered saline (TBS) solution with the detergent Tween 20, ABO Gdanska, Poland) buffer for 30 min. After blocking in 10% normal horse serum for 2 h RT (room temperature), the tissue section was incubated with primary antibodies: insulin (1:100 dilution, sc-8033; Santa Cruz Biotechnology, Dallas, TX, USA), glucagon (1:100 dilution, sc-514592; Santa Cruz Biotechnology, Dallas, TX, USA), CD45 (Novus Biologicals, Centennial, CO, USA, NB100-77417, 1:500), or MOMA (Novus Biologicals, Centennial, CO, USA; NB100-64946, 1:500) overnight at 4 °C. The tissue section used as the negative control was incubated with 10% normal horse serum overnight at 4 °C. Then, after washing, the sample sections were covered by AP- or HRP-conjugated secondary mouse antibody (Vector-ImmPRESS Reagent Kit, MP-5402, or Vector-ImmPRES Duet, MP-7724; Vector Laboratories, Newark, CA, USA) for 2 h RT. Following the three washing, the AP/DAB substrate (Vector-ImmPRES Duet, MP-7724, Vector Laboratories, Newark, CA, USA) was added to the tissue slides. When staining was well developed, slides were washed and then mounted.
Klak M., Wszoła M., Berman A., Filip A., Kosowska A., Olkowska-Truchanowicz J., Rachalewski M., Tymicki G., Bryniarski T., Kołodziejska M., Dobrzański T., Ujazdowska D., Wejman J., Uhrynowska-Tyszkiewicz I, & Kamiński A. (2023). Bioprinted 3D Bionic Scaffolds with Pancreatic Islets as a New Therapy for Type 1 Diabetes—Analysis of the Results of Preclinical Studies on a Mouse Model. Journal of Functional Biomaterials, 14(7), 371.
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5

Histological Analysis of Liver, Adipose, and Pancreas

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A piece of liver, epididymal white adipose tissue (eWAT) and pancreas were fixed in 10% formalin overnight, processed and embedded in paraffin wax. Approximately 4μm thick sections of tissues were deparaffinized and then stained with hematoxylin and eosin (H&E) or used for immunohistochemistry. Pancreatic sections were incubated with insulin (sc-8033, Santa Cruz Biotechnology Inc.) or glucagon (sc-514592, Santa Cruz Biotechnology Inc.) antibody overnight at 4°C and then incubated with horseradish peroxidase–conjugated secondary antibody. Immunoreactivities were detected with 3, 3′ diaminobenzidine tetrahydrochloride substrate (Vector Laboratories, Burlingame, CA) and counterstained with hematoxylin. Slides were examined under the Zeiss light microscope and images were taken at x20 magnification.
Tran M., Mostofa M.G., Picard M., Wu J., Wang L, & Shin D.J. (2023). SerpinA3N deficiency attenuates steatosis and enhances insulin signaling in male mice. The Journal of endocrinology, 256(3), e220073.
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6

Immunofluorescence Staining of Pancreatic Cells

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Cells were fixed in 4% paraformaldehyde for 15 min at room temperature (RT) and washed three times with PBS, followed by blocking with 5% normal goat serum for 1 h and incubation with primary antibodies at room temperature (1 h). The cells were then washed three times with PBS, and incubated with secondary antibody for 30 min. The following primary antibodies were used: a-Amylase (sc‐25562, rabbit polyclonal; Santa Cruz Biotechnology, dilution 1:100), cytokeratin 19 (A53-B/A2: sc-6278, mouse monoclonal; Santa Cruz Biotechnology, dilution 1:100), insulin (sc‐8033, mouse monoclonal; Santa Cruz Biotechnology, dilution 1:100), and vimentin (D21H3: #5741, mouse monoclonal; Cell signaling technology, dilution 1:100). The cells were stained using Alexa Flour-conjugated secondary antibodies from Invitrogen.
Lee H.S., Kim E., Lee J., Park S.J., Hwang H.K., Park C.H., Jo S.Y., Kang C.M., Hong S.M., Kang H., Jo J.H., Cho I.R., Chung M.J., Park J.Y., Park S.W., Song S.Y., Han J.M., Kim S, & Bang S. (2021). Profiling of conditionally reprogrammed cell lines for in vitro chemotherapy response prediction of pancreatic cancer. EBioMedicine, 65, 103218.
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7

Insulin Immunohistochemistry Protocol

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Slides were dewaxed and hydrated and then underwent antigen retrieval by high-temperature induced epitope retrieval (HIER) in citrate buffer (0.1 M, pH 6.0) in a high-pressure cooker for 3 minutes. Then the endogenous peroxidases were blocked by 3% H2O2 for 10 minutes. The mouse monoclonal primary antibody of insulin (SC-8033, Santa Cruz Biotechnology) with a dilution of 1:100 were incubated with the slides for one hour. The secondary antibody, MaxVisionTM HRP-Polymer anti-Mouse antibody (KIT-5001, Maxim Biotechnology), was incubated for 20 minutes. Then DAB was incubated for 5 minutes. Between each of the above steps, the slides were washed with PBS (0.1M, PH 7.2) 3 times for 3 minutes. After DAB coloration, the slides were counterstained with haematoxylin, dehydrated, and mounted with resinene.
Duan J., Yang M., Liu Y., Xiao S, & Zhang X. (2022). Curcumin protects islet beta cells from streptozotocin‑induced type 2 diabetes mellitus injury via its antioxidative effects. Endokrynologia Polska, 73(6).
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