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Immune precipitation assay buffer

Manufactured by Keygen Biotech
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Sourced in China

The Immune precipitation assay buffer is a laboratory reagent designed to facilitate the precipitation and isolation of antigen-antibody complexes during immunoprecipitation experiments. This buffer aids in the capture and purification of specific proteins or protein complexes from complex biological samples.

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Lab products found in correlation

Bca protein assay kit, by Keygen Biotech (3 mentions) Polyvinylidene difluoride membrane, by Pall Corporation (3 mentions) Phenylmethylsulfonyl fluoride, by Keygen Biotech (2 mentions) Ecl western blotting kit, by Advansta (2 mentions) Phosphatase inhibitor cocktail, by Keygen Biotech (2 mentions) Nuclear and cytoplasmic protein extraction kit, by Thermo Fisher Scientific (1 mentions)

3 protocols using immune precipitation assay buffer

1

Protein Extraction and Western Blot

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Cells were harvested and lysed in an immune precipitation assay buffer (KeyGen Biotech Co Ltd) supplemented with 1 mM phenylmethylsulfonyl fluoride (KeyGen Biotech Co Ltd) and 1 mM of a phosphatase inhibitor cocktail (KeyGen Biotech Co Ltd). The protein concentration was determined using a BCA protein assay kit (KeyGen Biotech Co Ltd). Same amounts of protein samples were separated in 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis gels and then transferred to a polyvinylidene difluoride membrane (Pall Corporation, Port Washington, NY). The protein band intensities were evaluated using ECL Western blot analysis kit (Advansta, Menlo Park, CA) and were normalized to those of β-actin.
Dong C., Liu Y., Jiang K., Wang H., Qu W., Zhang C., Liang R., Gao Z., Zhao B., Miao Q., Shao S, & Wang L. (2018). The Nogo-B receptor promotes human hepatocellular carcinoma cell growth via the Akt signal pathway. Journal of cellular biochemistry, 119(9), 7738-7746.
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2

Protein Extraction and Localization Analysis

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Cells were harvested and lysed in immune precipitation assay buffer (KeyGen Biotech Co., Ltd, Nanjing, China) supplemented with 1 mM phenylmethylsulfonyl floride (KeyGen Biotech Co., Ltd, Nanjing, China) and 1 mM phosphatase inhibitor cocktail (KeyGen Biotech Co., Ltd) for extracting whole cell protein samples. For detecting the cellular localization of total MDM2 and phos-MDM2, nuclear and cytoplasmic fractions were isolated using the Nuclear and Cytoplasmic Protein Extraction kit (Thermo Fisher Scientific) according to the instructions of the manufacturer. Protein concentration was determined using a BCA protein assay kit (KeyGen Biotech Co., Ltd). Same amount protein samples were separated on 10% SDS-PAGE gels and then transferred to a polyvinylidene difluoride membrane (Pall Corporation, Port Washington, NY, USA). The protein band intensities were evaluated using ECL western blotting kit (Advansta, Menlo Park, CA, USA) and were normalized to those of β-actin. All experiments were performed at least three times.
Dong C., Zhao B., Long F., Liu Y., Liu Z., Li S., Yang X., Sun D., Wang H., Liu Q., Liang R., Li Y., Gao Z., Shao S., Miao Q.R, & Wang L. (2016). Nogo-B receptor promotes the chemoresistance of human hepatocellular carcinoma via the ubiquitination of p53 protein. Oncotarget, 7(8), 8850-8865.
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3

Whole Cell Protein Extraction and Western Blot

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Cells were harvested and lysed in immune precipitation assay buffer (KeyGen Biotech Co., Ltd, Nanjing, China) supplemented with 1 mM phenylmethylsulfonyl fluoride (KeyGen Biotech Co., Ltd, Nanjing, China) and 1 mM phosphatase inhibitor cocktail (KeyGen Biotech Co., Ltd) for extracting whole cell protein samples. Protein concentration was determined using a bicinchoninic acid (BCA) protein assay kit (KeyGen Biotech Co., Ltd). Equal amounts of protein samples were separated on 10% SDS-PAGE gels and then transferred onto polyvinylidene difluoride membranes (Pall Corporation, Port Washington, NY, USA). The protein band intensities were evaluated using an electrochemiluminescence (ECL) western blotting kit (Advansta, Menlo Park, CA, USA) and were normalized to GAPDH or β-tubulin. All experiments were performed at least three times.
Mao J., Tian Y., Wang C., Jiang K., Li R., Yao Y., Zhang R., Sun D., Liang R., Gao Z., Wang Q, & Wang L. (2019). CBX2 Regulates Proliferation and Apoptosis via the Phosphorylation of YAP in Hepatocellular Carcinoma. Journal of Cancer, 10(12), 2706-2719.
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