Anti ha beads

Manufactured by Smart-Lifesciences

Anti-HA beads are a type of affinity chromatography resin used for the purification of proteins tagged with the hemagglutinin (HA) epitope. They consist of an agarose matrix with covalently bound anti-HA antibodies, which can selectively bind and capture HA-tagged proteins from complex mixtures.

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Lab products found in correlation

Anti flag beads, by Smart-Lifesciences (1 mentions) Protease inhibitor, by Roche (1 mentions) Protease inhibitors, by Smart-Lifesciences (1 mentions)

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Anti-HA Affinity Beads (2 citations)

2 protocols using anti ha beads

Analyzing Protein Interactions and Modifications in Adipocytes

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The messenger RNA expression level was analyzed by quantitative RT‐PCR. Western blot was used to explore the protein levels. The primer sequences and antibodies used in this study were listed in Table S2 and Table S3, Supporting Information.
For IP assay, HEK293T cell extracts with ectopic expressing tagged plasmids were incubated with anti‐HA beads (Smart lifesciences, SA068001) or anti‐Flag beads (Smart lifesciences, SA042001). Immunoprecipitates were probed with indicated antibodies. To examine the interaction between HDAC1 or ZFP516 with PWWP2B in brown fat cells, lentiviral HA‐PWWP2B or HA‐ZFP516 was transduced to immortalized brown preadipocytes. HA antibody was used to precipitate the HA‐PWWP2B or HA‐ZFP516 protein complex from differentiated mature adipocytes followed by western blot. The HA‐PWW2B protein complex was subjected to LC‐MS/MS analysis (Shanghai Applied Protein Technology Co. Ltd). For the ubiquitination assay, HEK293T cells were treated with 5 µM MG132 for 12 h before harvesting the cells.

Yan L., Jin W., Zhao Q., Cui X., Shi T., Xu Y., Li F., Jin W., Zhang Z., Zhang Z., Tang Q, & Pan D. (2021). PWWP2B Fine‐Tunes Adipose Thermogenesis by Stabilizing HDACs in a NuRD Subcomplex. Advanced Science, 8(16), 2102060.

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Detecting SUMOylated AMPKα in HEK-293T Cells

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The V5-SUMO plasmid, HA-AMPKα plasmid, FLAG-SENP2 plasmid, and FLAG-UBC9 plasmid were transfected into HEK-293T cells. HEK-293T cells were lysed in lysis buffer containing 50 mM Tris–Hcl (pH 8.0), 250 mM NaCl, 0.01% Tween, 1% Triton X-100, 0.0125 g/ml N-ethylmaleimide, and protease inhibitors (Roche). After centrifugation, lysates were incubated with anti-HA beads (SMART LIFESCIENCES) and rotated at 4 °C for 2 h. The beads were then washed four times with washing buffer (50 mM Tris–Hcl [pH 8.0], 250 mM NaCl, 0.01% Tween, 0.0125 g/ml N-ethylmaleimide, and protease inhibitors). Finally, the immunocoprecipitates were subjected to Western blotting using V5 antibody to detect SUMOylated AMPKα.

Dou X., Zhou W.Y., Ding M., Ma Y.J., Yang Q.Q., Qian S.W., Tang Y., Tang Q.Q, & Liu Y. (2021). The protease SENP2 controls hepatic gluconeogenesis by regulating the SUMOylation of the fuel sensor AMPKα. The Journal of Biological Chemistry, 298(2), 101544.

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