Total protein from each group was fractionated by SDS-PAGE and transferred onto a nitrocellulose membrane. The membrane was blocked with 5% non-fat milk and incubated with primary antibodies against α2A-adrenoreceptor (1:200, sc1478, Santa Cruz), β2-adrenoreceptor (1:500, ab137494, Abcam), aggrecan (1:500, ab36861, Abcam), MMP-3 (1:200, sc6839, Santa Cruz), MMP-13 (1:300, sc30073, Santa Cruz), RANKL (1:300, sc7628, Santa Cruz), β-actin (1:1000, 3700, Cell Signalling Technology, USA), Phospho-ERK1/2 (Thr202/Tyr204) (1:800, 4370, Cell Signalling Technology), total-ERK1/2 (1:1000, 4695, Cell Signalling Technology), phospho-p38 (Thr180/Tyr182) (1:800, 4511, Cell Signalling Technology), total-p38 (1:1000, 9212, Cell Signalling Technology), phospho-JNK (1:800, 4688, Cell Signalling Technology), total-JNK (1:1000, 9252, Cell Signalling Technology), phospho-Akt (Thr308) (1:800, 4056, Cell Signalling Technology) and total-Akt (1:1000, 4691, Cell Signalling Technology). Signals were revealed by incubation with a horseradish peroxidase-conjugated secondary antibody (1:5000, Zhongshan Golden Bridge Biotechnology, China) and enhanced chemiluminescence detection22 (link)23 (link)24 (link)25 (link).
Sc7628
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Sc7628 is a laboratory equipment product offered by Santa Cruz Biotechnology. It is a multi-functional device designed for various applications in the field of biotechnology research. The core function of the Sc7628 is to perform specific tasks related to sample preparation, analysis, or experimental procedures. No further details about the intended use or capabilities of the Sc7628 can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Ab36861, by Abcam (2 mentions)
Sc1478, by Santa Cruz Biotechnology (2 mentions)
Ab137494, by Abcam (2 mentions)
Sc 6839, by Santa Cruz Biotechnology (2 mentions)
Sc 30073, by Santa Cruz Biotechnology (2 mentions)
Sc 30833, by Santa Cruz Biotechnology (2 mentions)
Sc 8468, by Santa Cruz Biotechnology (2 mentions)
Total p38, by Cell Signaling Technology (1 mentions)
Phospho jnk, by Cell Signaling Technology (1 mentions)
Total erk1 2, by Cell Signaling Technology (1 mentions)
Phospho p38 thr180 tyr182, by Cell Signaling Technology (1 mentions)
Total jnk, by Cell Signaling Technology (1 mentions)
Phospho akt thr308, by Cell Signaling Technology (1 mentions)
Total akt, by Cell Signaling Technology (1 mentions)
Phospho erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Zhongshan Golden Bridge Biotechnology (1 mentions)
4 protocols using sc7628
1
Western Blot Analysis of Signaling Pathways
2
Quantitative Analysis of Cartilage Receptors
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Tissue processing, section staining and counting of immune-positive cells were performed as reported previously21 (link)22 (link)23 (link)24 (link)25 (link). The primary antibodies were goat polyclonal α2A-adrenoreceptor (1:75; sc1478, Santa Cruz Biotechnology, Inc., USA), rabbit polyclonal β2-adrenoreceptor (1:100, ab137494; Abcam, Cambridge, United Kingdom), rabbit polyclonal aggrecan (1:200, ab36861, Abcam), goat polyclonal MMP-3 (1:50, sc6839, Santa Cruz), MMP-13 (1:50, sc30073, Santa Cruz), RANKL (1:50, sc7628, Santa Cruz). Six squares were applied at the quartering points of the central (each 0.15 mm × 0.15 mm) and posterior (each 0.2 mm × 0.2 mm) third of the condylar cartilage. Within the selected frames, the number of immune-positive cells and the percentage area of aggrecan-positive staining were determined. In the isotype control slides, isotype antibodies were substituted for the primary antibodies.
3
Histochemical Analysis of Bone Remodeling
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The specimens were demineralized in 10% ethylenediaminetetraacetic acid (EDTA) and processed in a conventional manner. Semi-serial sections (4 μm) were obtained in the mesiodistal direction, and 5 equidistant sections of each specimen were stained with hematoxylin and eosin (H&E) for histological and histometric analyses. Other sections were subjected to indirect immunoperoxidase staining with the following primary antibodies: anti-tartrate-resistant acid phosphatase (anti-TRAP).(SC-30833, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-receptor activator of nuclear factor kappa-B ligand (anti-RANKL) (SC-7628, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-osteoprotegerin (anti-OPG) (SC-8468, Santa Cruz Biotechnology, Santa Cruz, CA, US). The immunohistochemical processing followed the protocol described by Garcia et al., (2013) [31 (link)].
4
Histological Analysis of Alveolar Bone
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An overdose of thiopental (150 mg/kg of body weight; Cristalia, Itapira, SP, Brazil) was administered to euthanize the animals (7, 15, or 30 days after removing the ligature). The left mandibles were excised, fixed in 4% formaldehyde, and decalcified in 10% ethylenediaminetetraacetic acid solution. After tissue processing, the specimens were embedded in paraffin. Semi-serial sections cut in a disto-mesial direction, 5 µm in thickness, were obtained in a buccal-lingual sequence. The most central buccal-lingual sections of the FA of left mandibular first molars were used for histological, histomorphometric, and immunohistochemical analyses. They were either stained with hematoxylin, eosin, and phloxine for histological and histomorphometric analyses or subjected to an indirect immunoperoxidase method with the following primary antibodies: anti-tartrate-resistant acid phosphatase (TRAP) (SC30833, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-receptor activator of nuclear factor-κB ligand (RANKL) (SC7628, Santa Cruz Biotechnology) and anti-osteoprotegerin (OPG) (SC8468, Santa Cruz Biotechnology). The immunohistochemical protocol was previously described by Nunes et al. [20 (link)].
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