Sa 5704

Manufactured by Vector Laboratories

The SA-5704 is a laboratory instrument designed for the detection and analysis of specific proteins or molecular targets. The core function of the SA-5704 is to provide precise and reliable measurements of target analytes in samples.

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Lab products found in correlation

Ba 1000, by Vector Laboratories (3 mentions) Ab16669, by Abcam (3 mentions) Streptavidin horseradish peroxidase, by Vector Laboratories (3 mentions) Ab183685, by Abcam (3 mentions) Cytoseal 60 mounting medium, by Thermo Fisher Scientific (3 mentions) Biotin conjugated secondary antibody, by Vector Laboratories (3 mentions) Eclipse 80i microscope, by Nikon (3 mentions) 3 3 diaminobenzidine (dab), by Vector Laboratories (3 mentions) Ab15580, by Abcam (2 mentions) Decalcifying solution, by Merck Group (1 mentions) Anti opn, by Abcam (1 mentions) Anti dspp, by Santa Cruz Biotechnology (1 mentions) Anti ocn, by Santa Cruz Biotechnology (1 mentions) Ba 5000, by Vector Laboratories (1 mentions) Zli 9017, by ZSGB-BIO (1 mentions) Bx53 microscope, by Olympus (1 mentions) K3468, by Agilent Technologies (1 mentions) Biotin, by Vector Laboratories (1 mentions) Sp 2001, by Vector Laboratories (1 mentions) Alexa fluor 594 conjugated goat anti rabbit igg, by ZSGB-BIO (1 mentions)

7 protocols using sa 5704

Histological and Immunohistochemical Analysis of Dental Extracellular Matrix Proteins

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The harvested samples were fixed in 10% neutral buffered formalin (NBF, Leica Biosystem) at room temperature for 24 h. Afterward, decalcified in decalcifying solution (Sigma‐Aldrich) at room temperature for 7 days. The samples were paraffin‐embedded and sectioned into 5 μm thick sections. Deparaffinized sections were stained with Hematoxylin and Eosin (H&E) staining.
For immunohistochemistry, deparaffinized sections were rehydrated and subjected to antigen retrieval at 37°C for 30 min. The samples were protein blocked with a serum‐free blocking solution at room temperature for 1 h, then, incubated with anti‐OPN (1:100 dilution, Abcam, MA, USA), anti‐OCN (1:50 dilution, Santa Cruz, CA, USA), anti‐DSPP (1:50 dilution, Santa Cruz, CA, USA), and ani‐DMP‐1 (1:500 dilution, Santa Cruz, CA, USA) primary antibodies at 4°C overnight. The samples were incubated with the secondary biotinylated anti‐goat antibody (BA‐5000, Vector Laboratories) for 30 min. Streptavidin‐conjugated horseradish peroxidase (SA‐5704, Vector Laboratories) was added for 30 min, and the samples were stained with 3,3′‐Diaminobenzidine (DAB). Then, Gill's hematoxylin was used to counterstain the cell nuclei. The stained sections were visualized under the light microscope and the positive areas of OPN, OCN, DSPP, and DMP‐1 were measured using Image J software.

Kim D., Lee H., Lee G., Hoang T., Kim H, & Kim G.H. (2022). Fabrication of bone‐derived decellularized extracellular matrix/ceramic‐based biocomposites and their osteo/odontogenic differentiation ability for dentin regeneration. Bioengineering & Translational Medicine, 7(3), e10317.

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Histological and Immunohistochemical Analysis of Ovarian Cancer

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Tissue specimens of OC patients and mice were fixed in 4% paraformaldehyde, dehydrated in ethanol and then paraffin-embedded. Consecutive sections (6 μm) were performed using microtome, and then deparaffinized in xylene. Tissue sections were stained with hematoxylin (ZLI-9610, Origene) and eosin (ZLI-9613, Origene) (HE) for morphological observations. For IHC staining, 3% H₂O₂ was used for antigen retrieval (C1031, Solarbio) and removing peroxidase. 5% goat serum was used for blocking tissue sections before incubated with primary antibodies, biotin-conjugated secondary antibodies (BA-1000, Vector Laboratories), and streptavidin-HRP (SA-5704, Vector Laboratories). The sections were visualized using 3,3'-diaminobenzidine (DAB) substrate (ZLI-9017, Zsgb-Bio), then images were taken using an Olympus BX53 microscope. The degree of immunostaining was measured by H score, and the final score was calculated by multiplying positively stained area (P) with staining intensity (I). The degrees for P were scored as 0-4: 0, < 5%; 1, 5%~25%; 2, 25%~50%; 3, 50%~75%; 4, 75%~100%. The degrees for I were scored as 0 - 3: 0, none; 1, weak; 2, moderate; 3, strong. The antibodies used in IHC staining were listed in Supplementary Table S2.

Qin X., Li J., Wang S., Lv J., Luan F., Liu Y., Chen Y., Chen X., Zhao Y., Zhu J., Piao Y., Zhang W., Shi Y., Xiang R., Qu P, & Wang L. (2021). Serotonin/HTR1E signaling blocks chronic stress-promoted progression of ovarian cancer. Theranostics, 11(14), 6950-6965.

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Immunohistochemistry of Mouse Tissues

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Mouse tissues were fixed in 10% buffer formalin solution overnight and then transferred to 70% ethanol for paraffin-embedded sections. Tissue sections were deparaffinized, rehydrated and boiled for 45 min in 10 mM pH 6 Citrate buffer. Slides were blocked in 10% goat serum for an hour and then incubated at 4˚C overnight with primary antibody against Ki67 (1:200, Ab15580, Abcam), active Caspase-3 (1:300, #9661, Cell Signaling), CD3 (1:100, Ab16669, Abcam), CD4 (1:1,000, Ab183685, Abcam) and CD8 (1:100, 14–0808-82, Invitrogen). The following day, tissue sections were incubated with biotin-conjugated secondary antibody for 15min (Vector Laboratories), 3% hydrogen peroxide for 5 min, horseradish peroxidase streptavidin for 15 min (SA-5704, Vector Laboratories) and developed by 3,3-diaminobenzidine (Vector Laboratories) followed by hematoxylin staining (3536–16, Ricca). Sections were then dehydrated, mounted in Cytoseal 60 mounting medium (8310, Thermo Scientific) and analyzed using Nikon Eclipse 80i microscope. For quantification of IHC, at least 10 images containing a minimum of 100 cells were analyzed at 60X magnification for each genotype.

Poillet-Perez L., Xie X., Zhan L., Yang Y., Sharp D.W., Hu Z.S., Su X., Maganti A., Jiang C., Lu W., Zheng H., Bosenberg M.W., Mehnert J.M., Guo J.Y., Lattime E., Rabinowitz J.D, & White E. (2018). Autophagy maintains tumor growth through circulating arginine. Nature, 563(7732), 569-573.

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Immunohistochemical Detection of Junin Virus

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Necropsy was performed on all animals, and representative tissue samples were collected. The tissues were stored for at least 21 d in 10% neutral buffered formalin in the BSL-4, and the formalin was changed prior to removal from the BSL-4. Formalin-fixed tissues were then processed in the BSL-2 and embedded in paraffin wax. Embedded tissue blocks were sectioned at 4 μm and dried at 60 °C. The sections were deparaffinized and rehydrated in xylene, graded ethanol, and deionized water. Antigen retrieval incubation was performed in a 10 mM, pH 6 citrate buffer for 20 min at 95 °C, then allowed to cool at room temperature for 20 min. The sections were then quenched in a solution of 3% hydrogen peroxide for 10 min before being processed for IHC analysis using the Thermo Scientific Lab Vision Autostainer 360. Avidin D and biotin (Vector Laboratories; SP-2001) were then applied for 15 min to block endogenous biotin reactivity. Specific anti-JUNV immunoreactivity was detected using an anti-JUNV primary antibody at 1:1,000 dilution for 1 h (LifeSpan Bio; LS-C500445). The secondary antibody used was biotinylated goat anti-rabbit IgG (Vector Laboratories; BA-1000) at 1:200 for 30 min, followed by streptavidin-HRP (Vector Laboratories; SA-5704) for 30 min. Slides were developed with DAB chromagen (Dako; K3468) for 5 min and then counterstained with hematoxylin for 1 min.

Zeitlin L., Cross R.W., Geisbert J.B., Borisevich V., Agans K.N., Prasad A.N., Enterlein S., Aman M.J., Bornholdt Z.A., Brennan M.B., Campbell L., Kim D., Mlakar N., Moyer C.L., Pauly M.H., Shestowsky W., Whaley K.J., Fenton K.A, & Geisbert T.W. (2021). Therapy for Argentine hemorrhagic fever in nonhuman primates with a humanized monoclonal antibody. Proceedings of the National Academy of Sciences of the United States of America, 118(11), e2023332118.

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Immunohistochemistry and Immunofluorescence Protocols

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Tissues were fixed with 4% paraformaldehyde for at least 24 hours. For HE staining, tissues were embedded in paraffin and sectioned into 4-μm intervals (Leica). For immunohistochemistry (IHC), tissue sections were deparaffinized, rehydrated, and permeated using Triton X 100 (T8200, Solarbio) and followed by antigen retrieval using EDTA Antigen Retrieval solution (c1034, Solarbio). The primary antibodies used for IHC and IF were the same as WB (dilution 1:200). Biotinylated goat anti-rabbit IgG antibody (BA-1000, Vector Laboratories), Streptavidin, Horseradish Peroxidase, R.T.U. (SA-5704, Vector Laboratories), and DAB kit (ZLI-9017, Zsgb-Bio) were used for IHC. Alexa Fluor 488—Conjugated Goat anti-Mouse IgG (ZF0512, Zsgb-Bio), Alexa Fluor 594—Conjugated Goat anti-Rabbit IgG (ZF0516, Zsgb-Bio) and DAPI (D9542, Sigma‐Aldrich) were used for IF. Fluoro-Gel Mounting Medium with TES Buffer (17985–30, Electron Microscopy Sciences) was used for protecting the fluorescence.

Lv Y., Wang X., Li X., Xu G., Bai Y., Wu J., Piao Y., Shi Y., Xiang R, & Wang L. (2020). Nucleotide de novo synthesis increases breast cancer stemness and metastasis via cGMP-PKG-MAPK signaling pathway. PLoS Biology, 18(11), e3000872.

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Immunohistochemical Analysis of Immune Markers in Mouse Tissues

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Mouse tissues were fixed in 10% buffer formalin solution overnight and then transferred to 70% ethanol for paraffin-embedded sections. Tissue sections were deparaffinized, rehydrated and boiled for 45 min in 10 mM (pH 6) citrate buffer. Slides were blocked in 10% goat serum for 1 h and then incubated at 4 °C overnight with primary antibody against CD3 (1:100 dilution, Ab16669, Abcam), CD4 (1:1,000 dilution, Ab183685, Abcam), CD8 (1:100 dilution, 14–0808-82, Invitrogen), Foxp3 (1:75 dilution, 14–5773-80, Invitrogen), FIP200 (Proteintech, 17250–1-AP, 1:100 dilution) and P62 (Enzo, P9860, 1:1,000 dilution). The following day, tissue sections were incubated with biotin-conjugated secondary antibody for 15 min (Vector Laboratories), 3% hydrogen peroxide for 5 min, horseradish peroxidase streptavidin for 15 min (SA-5704, Vector Laboratories) and developed by 3,3-diaminobenzidine (Vector Laboratories) followed by hematoxylin staining (3536–16, Ricca). Sections were then dehydrated, mounted in Cytoseal 60 mounting medium (8310, Thermo Fisher Scientific) and analyzed using a Nikon Eclipse 80i microscope. For quantification of IHC, at least 15 images were analyzed at ×60 magnification for each genotype.

Poillet-Perez L., Sharp D.W., Yang Y., Laddha S.V., Ibrahim M., Bommareddy P.K., Sherrie Hu Z., Vieth J., Haas M., Bosenberg M.W., Rabinowitz J.D., Cao J., Guan J.L., Ganesan S., Chan C.S., Mehnert J.M., Lattime E.C, & White E. (2020). Autophagy promotes growth of tumors with high mutational burden by inhibiting a T-cell immune response. Nature cancer, 1(9), 923-934.

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Immunohistochemistry of Mouse Tissues

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