This is a knockout-validated antibody summary, based on the publication "Sc65-Null Mice Provide Evidence for a Novel Endoplasmic Reticulum Complex Regulating Collagen Lysyl Hydroxylation", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Western blot confirmation of SC65 protein (arrow) loss in multiple Sc65KO tissues from 3 day-old mice compared to WT controls (lower panel—Cal = calvaria, Kid = Kidney). From [1].

Rabbit polyclonal

Company: ProteinTech

Antibody: Sc65

Catalog number: 15288-1-AP

Summary: Rabbit polyclonal against SC65 fusion protein. Reacts with human, mouse, and rat and it is recommended for western blot, immunoprecipitation and ELISA.

Western blot | Immunohistochemistry | Immunocytochemistry

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Figure 2. Immunohistochemistry detection of SC65 in adult femur section from a WT mouse showing specific intracellular staining in bone forming cells (osteoblasts) aligned on the surface of a bone trabecula. Sc65 expression is lost in a similar section from a Sc65KO mouse. Scale bars = 100μM (10x) or 20μM (63x). From [1].

WT and Sc65KO mouse tissues and primary cell cultures.

WB: cells were lysed into RIPA buffer (50mM Tris‐HCl pH 7.5, 150 mM NaCl, 0.1% SDS, 1 mM EDTA, 0.5% sodium deoxycholate, and 1% Triton X‐100) containing a cocktail of protease inhibitors including EDTA (cAMRESCO, Solon, OH, USA).

IHC: bone and skin sections were mounted on slides.

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Figure 3. Sc65 is expressed in WT dermal fibroblasts; its expression is lost in skin sections from Sc65KO mice. From [1].

IC: de-paraffinized mouse skin sections.

WB: 5% milk for 30min

IHC: blocking buffer supplied in the CTS008 Anti-Goat HRP-DAB Cell and Tissue Staining Kit (R&D Systems).

IC: normal serum blocking buffer (3% goat serum, 1 mg/ml BSA, 0.1% Triton-X) for 1 hour

IHC: overnight incubation.

IC: overnight incubation.

WB: goat anti‐rabbit IRDye 680LT (LICOR Biosciences, Lincoln, NE, USA).

IHC: secondary antibody for 30 minutes.

IC: AlexaFluor 488 goat anti-rabbit secondary antibody (Life Technologies) for 30 minutes.

WB: using a LICOR Odyssey instrument.

IHC: Zeiss AxioImager equipped with Zeiss AxioCam MRc5 digital camera and Zeiss Axiovision software.

IC: Zeiss AxioImager scope.

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.