Mouse anti rhodopsin

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Mouse anti-rhodopsin is a monoclonal antibody that specifically recognizes the rhodopsin protein. Rhodopsin is a light-sensitive G-protein-coupled receptor found in the retinal photoreceptor cells of vertebrates, where it plays a crucial role in the visual transduction process. The mouse anti-rhodopsin antibody can be used for the detection and characterization of rhodopsin in various applications, such as immunohistochemistry, Western blotting, and flow cytometry.

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Rhodopsin (4 citations)

3 protocols using mouse anti rhodopsin

Western Blot Analysis of SMN Protein

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Cell pellets were collected and lysed in lysis buffer with protease inhibitor cocktail (Sigma-Aldrich). The particulate fraction was removed by centrifugation. Proteins (20-40 μg) were separated on 10% SDS–PAGE and subjected to immunoblotting analysis (Li et al, 2009 (link)). Primary antibodies used were rabbit anti-actin (1:1,000; Sigma-Aldrich), rabbit anti-GAPDH (1:2,000; ExCell Bio), mouse anti-SMN (1:1,000; Abnova), mouse anti-Myc (1:2,000; Cell Signaling Technology), rabbit anti-ubiquitin (1:1,000; Cell Signaling Technology), mouse anti-FLAG (1:1,000; Sigma-Aldrich), rabbit anti-HA (1:1,000; Cell Signaling Technology), and mouse anti-rhodopsin (Huang et al, 2006 (link)) (1:1,000; Santa Cruz). Horseradish peroxidase-conjugated secondary antibodies were detected with Western Lighting Chemiluminescence Reagent Plus (Pierce). For quantifying SMN proteins, SMN band intensities were normalized with actin or GAPDH and compared between different groups using ImageJ software.

Wang Y., Xu C., Ma L., Mou Y., Zhang B., Zhou S., Tian Y., Trinh J., Zhang X, & Li X.J. (2019). Drug screening with human SMN2 reporter identifies SMN protein stabilizers to correct SMA pathology. Life Science Alliance, 2(2), e201800268.

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Immunofluorescence Characterization of Cultured Cells

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Cultures were fixed in 4% paraformaldehyde (PFA), and after PBS rinse, non-specific binding was inhibited by incubation in milk powder block solution (5% dried milk powder and 0.5% Triton X-100 in PBS) for 1–2 hours (at room temperature). Cells were incubated in primary antibodies overnight at 4°C, washed in PBS and incubated with appropriate fluorescent conjugated secondary antibodies (Invitrogen) or streptavidin conjugate (Invitrogen) for 1 hour along with DAPI to counterstain nuclei. The coverslips or sections were washed and mounted for microscopy. Primary antibodies were as follows: rabbit anti-Pax6 (1:500, PRB-278P, Covance, Princeton, NJ, USA), goat anti-Otx2-biotin (1:500, BAF1979, R&D Systems, Minneapolis, MN, USA), goat anti-Sox2 (1:100, sc17320, Santa Cruz), goat anti-Lhx2 (1:250, sc-19344, Santa Cruz), Tuj1 (1:3,000, Covance), Tbr2 (1:500, ab23345, Abcam), goat anti-GIPC1 (1:100, Santa Cruz), goat anti-Brn3 (1:500, sc-6026, Santa Cruz), rabbit anti-Recoverin (1:1000, Chemicon), Lin28 (1:100, ab46020, Abcam), chicken anti-GFP (1:250, Abcam), mouse anti-Rhodopsin (1:100), goat anti-S-Opsin (1:150, Santa Cruz), goat anti-Oct3/4 (1:200, sc-8628, Santa Cruz), rabbit Nanog (1:250, ab80892, Abcam).

La Torre A., Hoshino A., Cavanaugh C., Ware C.B, & Reh T.A. (2015). The GIPC1-Akt1 pathway is required for the specification of the eye field in mouse embryonic stem cells. Stem cells (Dayton, Ohio), 33(9), 2674-2685.

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Western Blot Analysis of Retinal Proteins

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Proteins were extracted using the radio-immunoprecipitation assay (RIPA) buffer (RiboBio; China) containing a protease inhibitor cocktail (Sigma, USA). Lysate proteins (50 μg protein per gel lane) were separated by 15% SDS-PAGE and transferred onto polyvinylidene fluoride (PVDF) membranes (280 mA for 55 min). The membranes were blocked with 5% skimmed milk in Tris buffered saline with 0.1% Tween-20 (TBS-T) for 1 h at 37 °C, then incubated with primary antibodies(mouse anti-rhodopsin, 1:500; rabbit anti-CRX, 1:500, Santa Cruz, sc-30150; rabbit anti-S-opin, 1:20, Abcam, ab81017) overnight at 4 °C. Immunoblotting with a mouse α-tubulin antibody (1:10000, Proteintech, 66031-1-Ig) was used as the gel loading control. After several washes, the membranes were incubated with horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Multi Science, China) or HRP-goat anti-rabbit IgG (Multi Science, China). The bands were semiquantified by densitometry using Bio-Rad imaging software.

Ji H.P., Xiong Y., Song W.T., Zhang E.D., Gao Z.L., Yao F., Su T., Zhou R.R, & Xia X.B. (2017). MicroRNA-28 potentially regulates the photoreceptor lineage commitment of Müller glia-derived progenitors. Scientific Reports, 7, 11374.

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