A2220 1ml

Manufactured by Merck Group

The A2220-1ML is a laboratory product offered by Merck Group. It serves as a core functional component for various scientific applications. The product specification and details are available upon request.

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Lab products found in correlation

Protein a g plus agarose bead, by Santa Cruz Biotechnology (2 mentions) P8345 5ml, by Merck Group (2 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Anti myc antibody, by Merck Group (1 mentions) F1804 200ug, by Merck Group (1 mentions) A7470 1 ml, by Merck Group (1 mentions) Hek293t cell, by American Type Culture Collection (1 mentions) Usinglipofectamine 2000, by Thermo Fisher Scientific (1 mentions)

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A2220 (41 citations)

4 protocols using a2220 1ml

Co-Immunoprecipitation of Flag- and GFP-Tagged Proteins

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For co-immunoprecipitation, HEK293 cells in 10-cm culture plates were transiently transfected with 6 μg of the indicated plasmids, cultured for 48 hours after transfection, and lysed in lysis buffer (150 mM NaCl, 20 mM Tris-HCl, 1.5 mM MgCl₂, 1% NP-40, 15% glycerol, 2 mM EDTA) with protease inhibitor cocktails (P8345-5ML, Sigma-Aldrich) at a dilution of 1:100. After precleaning cell lysates with protein A/G plus-agarose beads (sc-2003, Santa Cruz) for 1 hour and blocking anti-Flag (A2220-1ML, Sigma-Aldrich)/anti-GFP antibody-conjugated agarose beads with 2.5% albumin/bovine (94349-60-7, Acros Organics, Thermo Fisher, Waltham, MA) for 1 hour, cell lysates were added to anti-FLAG/anti-GFP antibody-conjugated agarose beads and rotated at 4° C for 1.5 hours. The beads were washed using lysis buffer four times. Immunoprecipitates and total cell lysates were boiled in SDS loading buffer for 10 minutes and then subjected to Western blot analysis using anti-Flag antibody or anti-GFP antibody.

Zang Y., Pascal L.E., Zhou Y., Qiu X., Wei L., Ai J., Nelson J.B., Zhong M., Xue B., Wang S., Yang D., Lan L., Shan Y, & Wang Z. (2017). ELL2 regulates DNA non-homologous end joining (NHEJ) repair in prostate cancer cells. Cancer letters, 415, 198-207.

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Antibody Detection Techniques

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6x-His epitope tag antibody (His.H8) was purchased from Invitrogen. Monoclonal Flag M2 antibody (F1804-50UG) and anti-Flag M2 affinity gel (A2220-1ML) were purchased from Sigma-Aldrich. Goat anti-mouse light chain–only HRP was purchased from Jackson Labs (115-035-174).

Rayaprolu V., Miettinen H.M., Baker W.D., Young V.C., Fisher M., Mueller G., Rankin W.O., Kelley J.T., Ratzan W.J., Leong L.M., Davisson J.A., Baker B.J, & Kohout S.C. (2024). Hydrophobic residues in S1 modulate enzymatic function and voltage sensing in voltage-sensing phosphatase. The Journal of General Physiology, 156(7), e202313467.

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Co-Immunoprecipitation Assay for Protein Interactions

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For co-immunoprecipitation, HEK 293 cells in 10-cm culture plates were transiently transfected with 6 μg of the indicated plasmids, cultured for 48 hours after transfection, and lysed in lysis buffer (150 mM NaCl, 20 mM Tris-HCl, 1.5 mM MgCl₂, 1% NP-40, 15% glycerol, 2 mM EDTA) with protease inhibitor cocktails (P8345-5ML, Sigma) at a dilution of 1:100. After precleaning cell lysates with protein A/G plus-agarose beads (sc-2003, Santa Cruz) for 1 hour and blocking anti-Flag (A2220-1ML, Sigma)/anti-MYC antibody-conjugated agarose beads (A7470-1ML, Sigma) with 2.5% albumin/bovine (94349-60-7, Acros organics) for 1 hour, cell lysates were added to anti-FLAG/anti-MYC antibody-conjugated agarose beads and rotated at 4°C for 1.5 hours. The beads were washed using lysis buffer four times. Immunoprecipitates and total cell lysates were boiled in SDS loading buffer for 10 minutes and then subjected to Western blot analysis using anti-Flag antibody (F1804-200UG, Sigma) and anti-MYC antibody (06-549, Sigma).

Qiu X., Pascal L.E., Song Q., Zang Y., Ai J., O’Malley K.J., Nelson J.B, & Wang Z. (2017). Physical and Functional Interactions between ELL2 and RB in the Suppression of Prostate Cancer Cell Proliferation, Migration, and Invasion. Neoplasia (New York, N.Y.), 19(3), 207-215.

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Hypoxia-Inducible Factor Regulation

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HEK293T cells (ATCC; no perceived need for authentication or mycoplasma
testing) were cotransfected with the indicated expression plasmids using
Lipofectamine 2000 (Invitrogen) following manufacturer’s instructions.
After 36 h, cells were treated with PT2399 (10 µM) at 37° C for
5 h, harvested for immunoprecipitation with anti-FLAG beads (A2220-1ML, Sigma)
and then subjected to western blot analysis. Plasmids laboratory database ID
#206 (HIF-1α) and #207 (HIF-2α) were used for in
vitro translation. Plasmids #930 (pcDNA3.1 Flag-HIF1β),
#931 (pcDNA3.1 Flag-HIF1β [F446L]), #932 (pLVX
HA-HIF-2α-IRES-zsGreen), and #933 (pLVX-HA-HIF-2α
[G323E]-IRES-zsGreen).

Chen W., Hill H., Christie A., Kim M.S., Holloman E., Pavia-Jimenez A., Homayoun F., Ma Y., Patel N., Yell P., Hao G., Yousuf Q., Joyce A., Pedrosa I., Geiger H., Zhang H., Chang J., Gardner K.H., Bruick R.K., Reeves C., Hwang T.H., Courtney K., Frenkel E., Sun X., Zojwalla N., Wong T., Rizzi J.P., Wallace E.M., Josey J.A., Xie Y., Xie X.J., Kapur P., McKay R.M, & Brugarolas J. (2016). Targeting Renal Cell Carcinoma with a HIF-2 antagonist. Nature, 539(7627), 112-117.

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