Sc 6486

Manufactured by Santa Cruz Biotechnology

Sc-6486 is a laboratory instrument designed for performing Western blot analysis. It is used to detect and quantify specific proteins in a sample. The device utilizes electrophoresis to separate proteins based on their molecular weight, and then transfers the separated proteins to a membrane for further analysis.

Automatically generated - may contain errors

Lab products found in correlation

4 protocols using sc 6486

Immunofluorescence Analysis of Cellular Markers

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence analysis was performed as previously described (Li et al., 2020 (link)). Briefly, free-floating sections were pre-incubated in blocking solution containing 5% normal donkey serum at room temperature for 1 h. Then primary antibodies were dissolved in diluent and incubated with sections overnight at 4°C. The following primary antibodies were used: TH (1:1,000, #AB9702, Millipore), TFE3 (1:500, #ab93808, Abcam), LC3 (1:100, #2775, Cell signaling Technology), Lamp1 (1:1,000, #1D4B-C, DSHB), CatD (1:200, #SC-6486, Santa Cruz Biotechnology), and p62 (1:1,000, #GP62-C, Progen Biotechnik). After three washes, the sections were then incubated with the secondary antibodies (Thermo Fisher), which were conjugated with Alexa 488, Alexa 555, or Alexa 647, at room temperature for 1 h. Finally, the sections were visualized under a confocal laser scanning microscope (LSM 780, Carl Zeiss) and the immunofluorescent images were quantified using ImageJ software.

He X., Xie Y., Zheng Q., Zhang Z., Ma S., Li J., Li M, & Huang Q. (2021). TFE3-Mediated Autophagy is Involved in Dopaminergic Neurodegeneration in Parkinson’s Disease. Frontiers in Cell and Developmental Biology, 9, 761773.

+ Open protocol

+ Expand

Analyzing Plac8 Protein Localization

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following antibodies were used: rabbit polyclonal antibody raised against the c-terminal 16 amino acids of murine Plac8 (Ledford et al., 2007 (link)) (Pocono Rabbit Farm & Laboratories, Inc.), rabbit polyclonal anti-β-Tubulin antibody (H-235, Santa Cruz), rat monoclonal anti-Lamp2 antibody (GL2A7, Abcam), mouse monoclonal anti-Rab7 antibody (R8779, Sigma), rabbit polyclonal anti-LC3B antibody (L7543, Sigma), mouse monoclonal anti-RhoA antibody (sc-418, Santa Cruz), goat polyclonal anti-Cathepsin D antibody (sc-6486, Santa Cruz), guinea pig polyclonal anti-p62 antibody (GP62-C, PROGEN), mouse monoclonal anti-3xFlag antibody and HRP-conjugated anti-3xFlag antibody (F2426 and A8592, Sigma), rabbit polyclonal anti-Rab5 antibody (ab18211, Abcam), rabbit polyclonal anti-Atg12 antibody (A8731, Sigma), EGFR antibody (sc 03).
The following compounds were used: bafilomycin A1 (Sigma B1793), chloroquine (Sigma 6628), cycloheximide (Sigma C7698, Lysosensor Yellow Blue DND-160 and DCF-DA (Molecular Probes, L-7545 and C6827, respectively).

Kinsey C., Balakrishnan V., O’Dell M.R., Huang J.L., Newman L., Whitney-Miller C.L., Hezel A.F, & Land H. (2014). Plac8 links oncogenic mutations to regulation of autophagy and is critical to pancreatic cancer progression. Cell reports, 7(4), 1143-1155.

+ Open protocol

+ Expand

Comprehensive Protein Detection Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary antibodies for western blotting (WB) and immunocytochemistry (ICC) were purchased and used at the indicated dilution; anti-LC3 (PM036, MBL; WB 1:3000; ICC 1:1000), anti-p62 (PM045, MBL; WB 1:5000), anti-Lamp1 (clone H4A3, sc-20011, Santa Cruz Biotechnology; ICC 1:1000), anti-EGFR (MI-12-1, MBL; WB 1:500), anti-cathepsin D (sc-6486, Santa Cruz Biotechnology; WB 1:1000), anti-EPG5 (sc-85198, Santa Cruz Biotechnology; WB 1:200), anti-GAPDH (MAb314, Chemicon; WB 1:5000), anti-α-tubulin (clone B-5-1-2, T5168, Sigma-Aldrich; WB 1:5000), anti-β-actin (clone 6D1, M177-3, MBL; WB 1:3000). BafA1 was used at 125 nM. PI comprised 10 μg/ml E64d and 10 μg/ml pepstatin A.

Hori I., Otomo T., Nakashima M., Miya F., Negishi Y., Shiraishi H., Nonoda Y., Magara S., Tohyama J., Okamoto N., Kumagai T., Shimoda K., Yukitake Y., Kajikawa D., Morio T., Hattori A., Nakagawa M., Ando N., Nishino I., Kato M., Tsunoda T., Saitsu H., Kanemura Y., Yamasaki M., Kosaki K., Matsumoto N., Yoshimori T, & Saitoh S. (2017). Defects in autophagosome-lysosome fusion underlie Vici syndrome, a neurodevelopmental disorder with multisystem involvement. Scientific Reports, 7, 3552.

+ Open protocol

+ Expand

Macrophage Lysosomal Dynamics Assessment

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human macrophages cultured in 24-well plates were treated as indicated above. In the last hour of infection, culture supernatants were replaced with medium that contained Lysotracker DND-99 Red (Life technologies) (100 nM). The slides were examined using a Zeiss Meta 510 laser confocal microscope with a plan-Apochromat 63× objective lens. A total of 10 fields containing 5–10 macrophages per field were examined in each experiment. The mean fluorescent intensity (MFI) for each macrophage was calculated using Image J software. Each cell from the image was selected and histogram analysis was performed. For immunostaining, mouse monoclonal antibodies for CD63 (sc-5275, Santa Cruz Biotechnology) and V1-ATPase H (sc-166227, santa Cruz Biotechnology) were visualized with anti-mouse-Alexafluor 588-conjugated secondary antibody. Goat polycolonal antibodies for active cathepsin D (sc-6486, Santa cruz) were visualized with anti-goat-Alexafluor 488-conjugated secondary antibody (Life technologies).

Jung J.Y, & Robinson C.M. (2014). IL-12 and IL-27 regulate the phagolysosomal pathway in mycobacteria-infected human macrophages. Cell Communication and Signaling : CCS, 12, 16.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!