Horseradish peroxidase hrp labeled secondary antibody

Manufactured by Elabscience

Horseradish peroxidase (HRP) labeled secondary antibody is a type of enzyme-linked antibody. It is used as a detection reagent in various immunoassay techniques, such as Western blotting, ELISA, and immunohistochemistry. The HRP enzyme catalyzes a colorimetric or chemiluminescent reaction, allowing for the visualization and quantification of target proteins or molecules.

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Lab products found in correlation

Ecl kit, by Elabscience (2 mentions) Tmem100, by OriGene (1 mentions) Bca kit, by Elabscience (1 mentions) Interleukin 6 (il 6), by Affinity Biosciences (1 mentions) Ripa lysis buffer, by Solarbio (1 mentions) Bca kit, by Solarbio (1 mentions)

2 protocols using horseradish peroxidase hrp labeled secondary antibody

Western Blot Analysis of TMEM100 and IL-6

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The tissue was lysed, and protein was extracted by radioimmunoprecipitation (RIPA) cleavage buffer (Solarbio, Beijing, China). The concentration of proteins was determined by a BCA kit (Elabscience). The separation gel and concentrated gel were prepared and sampled according to the determined concentration, then transferred to the polyvinylidene fluoride (PVDF) membrane after electrophoresis and sealed with 5% skimmed milk powder for 1 hour. Primary antibody was added to the PVDF membrane and incubated overnight at 4℃ (Table 1). The antibodies used were IL-6 (AffinityBiosciences), TMEM100 (Origene) and ß-actin (Elabscience). After overnight incubation, the PVDF membrane was incubated with horseradish peroxidase (HRP) labeled secondary antibody (Elabscience) for 1 hour, and then the luminescence was visualized by ECL kit (Elabscience). The obtained image was analyzed using a development device (Odyssey ® XF).

Table 1

Primary Antibodies and IgG Controls Used in This Study

Antibody	Host	Supplier/Catalog No.	Dilution
TMEM100	Mouse monoclonal	Origene/TA500532	1:100 (IHC), 1:500 (Wb)
IL-6	Rabbit polyclonal	Affinity/DF6087	1:1000 (Wb), 1:100 (IHC)
β-actin	Rabbit polyclonal	Elabscience/E-AB-20058	1:1000 (Wb)
IgG control	Mouse	Elabscience/E-AB-1001	1:2000 (Wb)
IgG control	Rabbit	Elabscience/E-AB-1003	1:5000 (Wb)
IgG control	Mouse	Elabscience/E-AB-1015	1:100 (IHC)
IgG control	Rabbit	Elabscience/E-AB-1014	1:100 (IHC)

Guo Z., Ma Y., Wang Y., Xiang H., Yang S.Y., Guo Z., Wang R., Chen W., Xing D., Chen B., Tao H, & Wu X. (2023). The Role of IL-6 and TMEM100 in Lumbar Discogenic Pain and the Mechanism of the Glycine-Serine-Threonine Metabolic Axis: A Metabolomic and Molecular Biology Study. Journal of Pain Research, 16, 437-461.

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Protein Extraction and Western Blot Analysis

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Proteins were extracted by lysing tissues with radioimmunoprecipitation (RIPA) lysis buffer (Solarbio, Beijing, China) containing 1 mM phenylmethanesulfonyl fluoride (PMSF). The concentration of extracted protein was determined with a bicinchoninic acid (BCA) kit (Solarbio, Beijing, China). Protein and loading buffer were mixed at a ratio of 4 : 1 (V/V) and boiled at 99°C for 10 min. Separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to PVDF membrane. PVDF membranes were blocked with 5% nonfat dry milk at room temperature. Membranes were then incubated with primary antibodies overnight at 4°C (Table 2). After overnight, the membrane was incubated with horseradish peroxidase (HRP)-labeled secondary antibody (Elabscience) for 1 hr, and then the ECL kit (Elabscience) was used for luminescence observation. The acquired images were analyzed using a developing instrument (Odyssey® XF).

Cui H., Guo Z., Guo Z., Fan Z., Shen N., Qi X., Ma Y., Zhu Y., Wu X., Chen B, & Xiang H. (2023). TMEM100 Regulates Neuropathic Pain by Reducing the Expression of Inflammatory Factors. Mediators of Inflammation, 2023, 9151967.

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