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Pmca2 pa1 915

Manufactured by Thermo Fisher Scientific
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PMCA2 (PA1-915) is a laboratory equipment product offered by Thermo Fisher Scientific. It is a plasma membrane calcium ATPase 2 (PMCA2) antibody that can be used in various research applications. The product's core function is to detect and analyze PMCA2 protein expression in biological samples.

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Lab products found in correlation

Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (2 mentions) Lsm 880, by Zeiss (2 mentions) 780 confocal microscope, by Zeiss (2 mentions) Lamp2 ab13524, by Abcam (2 mentions) Cathepsin b pa5 17007, by Thermo Fisher Scientific (2 mentions) Tfeb pa5 96632, by Thermo Fisher Scientific (2 mentions) Histoclear, by National Diagnostics (2 mentions) Specific secondary antibodies, by LI COR (2 mentions) Odyssey infrared imaging system, by LI COR (2 mentions) Wet wwestern blot transfer, by Bio-Rad (2 mentions) Pcna sc 25280, by Santa Cruz Biotechnology (1 mentions) Image lab software, by Bio-Rad (1 mentions) Sds page gel, by Thermo Fisher Scientific (1 mentions) Chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Hdac6 h 300, by Santa Cruz Biotechnology (1 mentions) Chemidoc xrs system, by Bio-Rad (1 mentions) Trans blot turbo system, by Bio-Rad (1 mentions) Sirt2, by Cell Signaling Technology (1 mentions)

5 protocols using pmca2 pa1 915

1

Immunofluorescence Staining of Cells and Tissues

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Cells were grown on coverslips, fixed in 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X100 for 10 min, washed three times with PBS and incubated with primary antibody overnight at 4 °C. The cells were washed three times with PBS and incubated with secondary antibody for 1 h at room temperature. After washing, coverslips were mounted using Prolong Gold antifade reagent with DAPI (Invitrogen). Paraffin-embedded tissue sections were cleared with histoclear (National Diagnostics) and graded alcohol using standard techniques. Antigen retrieval was performed using 7 mM citrate buffer, pH 6.0 under pressure. Sections were incubated with primary antibody overnight at 4 °C and with secondary antibody for 1 h at room temperature. Coverslips were mounted using Prolong Gold antifade reagent with DAPI (Invitrogen). All images were obtained using a Zeiss 780 confocal microscope and Zeiss LSM 880, and settings were adjusted to allow for detection of fine membrane structure. Primary antibodies were against: LAMP2 (ab13524) from Abcam (Cambridge, MA); PMCA2 (PA1-915) from Thermo Scientific (Waltham, MA); cathepsin B (PA5-17007) and TFEB (PA5-96632) from Invitrogen (Grand Island, NY); Phospho-STAT3 (9145), p21 (2947), p-Rb (8516), and NFAT (5861) from cell signaling (Danvers, MA); PCNA (sc-25280) from Santa Cruz (Dallas, TX).
Jeong J., Lee J., Talaia G., Kim W., Song J., Hong J., Yoo K., Gonzalez D.G., Athonvarangkul D., Shin J., Dann P., Haberman A.M., Kim L.K., Ferguson S.M., Choi J, & Wysolmerski J. (2024). Intracellular calcium links milk stasis to lysosome-dependent cell death during early mammary gland involution. Cellular and Molecular Life Sciences: CMLS, 81(1), 29.
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2

Western Blot Analysis of Protein Interactions

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Cell lysates and co-immunoprecipitated samples were separated on 4–12% SDS–PAGE gels (Thermo Fisher Scientific) and then transferred to membrane using Bio-Rad Trans-Blot Turbo system. Membranes were blocked; incubated with primary antibodies—acetylated α-tubulin (6-11B-1; Santa Cruz Biotechnology), ATAT1 (ab58742; Abcam), SIRT2 (12650; Cell Signalling Technology), HDAC6 (H-300; Santa Cruz Biotechnology), PMCA2 (PA1-915; Thermo Fisher Scientific), and NPC1 (NB400-148; Novus Bioscience); washed, incubated with appropriate HRP-conjugated secondary antibody and developed with chemiluminescent substrate (Thermo Fisher Scientific). Images were captured on a Bio-Rad ChemiDoc XRS+ system and quantified using ImageLab software (Bio-Rad). Membranes were then stripped and re-probed with primary antibodies.
Colaco A., Fernández-Suárez M.E., Shepherd D., Gal L., Bibi C., Chuartzman S., Diot A., Morten K., Eden E., Porter F.D., Poulton J., Platt N., Schuldiner M, & Platt F.M. (2020). Unbiased yeast screens identify cellular pathways affected in Niemann–Pick disease type C. Life Science Alliance, 3(7), e201800253.
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3

Immunoblotting of Protein Samples

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Protein samples were prepared from cells using standard methods, were subjected to SDS-PAGE and transferred to a nitrocellulose membrane by wet Wwestern blot transfer (Bio-Rad). The membrane was blocked in TBST buffer (TBS + 1% Tween) containing 5% milk for 1 hour at room temperature. The blocked membranes were incubated overnight at 4°C with specific primary antibodies (Odyssey blocking buffer, 927-40000). The membranes were washed 3 times with TBST buffer, and then incubated with specific secondary antibodies provided by LI-COR for 2 hours at room temperature. After 3 washes with TBST buffer, the membranes were analyzed using the ODYSSEY Infrared Imaging system (LI-COR). All immunoblot experiments were performed at least 3 times and representative blots are shown in the figures. Primary antibodies included those against: phospho-HER2 (thr1221/1222) (2243S), EGFR (4267S), AKT (4691S), phospho-AKT (4060S) from Cell Signaling (Danvers, MA); HER2 (sc-33684), phospho-EGFR (sc-12351), flotillin1 (sc-25506) from Santa Cruz (Dallas, Texas); PMCA2 (PA1-915) from Thermo Scientific (Waltham, MA).
Jeong J., Shin J.H., Li W., Hong J.Y., Lim J., Hwang J.Y., Chung J.J., Yan Q., Liu Y., Choi J, & Wysolmerski J. (2021). MAL2 mediates the formation of stable HER2 signaling complexes within lipid raft-rich membrane protrusions in breast cancer cells. Cell reports, 37(13), 110160.
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4

Immunoblotting of Protein Samples

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Protein samples were prepared from cells using standard methods, were subjected to SDS-PAGE and transferred to a nitrocellulose membrane by wet Wwestern blot transfer (Bio-Rad). The membrane was blocked in TBST buffer (TBS + 1% Tween) containing 5% milk for 1 hour at room temperature. The blocked membranes were incubated overnight at 4°C with specific primary antibodies (Odyssey blocking buffer, 927-40000). The membranes were washed 3 times with TBST buffer, and then incubated with specific secondary antibodies provided by LI-COR for 2 hours at room temperature. After 3 washes with TBST buffer, the membranes were analyzed using the ODYSSEY Infrared Imaging system (LI-COR). All immunoblot experiments were performed at least 3 times and representative blots are shown in the figures. Primary antibodies included those against: phospho-HER2 (thr1221/1222) (2243S), EGFR (4267S), AKT (4691S), phospho-AKT (4060S) from Cell Signaling (Danvers, MA); HER2 (sc-33684), phospho-EGFR (sc-12351), flotillin1 (sc-25506) from Santa Cruz (Dallas, Texas); PMCA2 (PA1-915) from Thermo Scientific (Waltham, MA).
Jeong J., Shin J.H., Li W., Hong J.Y., Lim J., Hwang J.Y., Chung J.J., Yan Q., Liu Y., Choi J, & Wysolmerski J. (2021). MAL2 mediates the formation of stable HER2 signaling complexes within lipid raft-rich membrane protrusions in breast cancer cells. Cell reports, 37(13), 110160.
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5

Immunofluorescence Staining of Cells and Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were grown on coverslips, fixed in 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X100 for 10 mins, washed 3 times with PBS and incubated with primary antibody overnight at 4°C. The cells were washed 3 times with PBS and incubated with secondary antibody for 1 hour at room temperature. After washing, coverslips were mounted using Prolong Gold antifade reagent with DAPI (Invitrogen). Paraffin-embedded tissue sections were cleared with histoclear (National Diagnostics) and graded alcohol using standard techniques. Antigen retrieval was performed using 7mM citrate buffer, pH 6.0 under pressure. Sections were incubated with primary antibody overnight at 4°C and with secondary antibody for 1 hour at room temperature. Coverslips were mounted using Prolong Gold antifade reagent with DAPI (Invitrogen). All images were obtained using a Zeiss 780 confocal microscope and Zeiss LSM 880, and settings were adjusted to allow for detection of fine membrane structure. Primary antibodies were against: LAMP2 (ab13524) from Abcam (Cambridge, MA); PMCA2 (PA1-915) from Thermo Scientific (Waltham, MA); cathepsin B (PA5-17007) and TFEB (PA5-96632) from Invitrogen (Grand Island, NY); Phospho-STAT3 (9145), p21 (2947), p-Rb (8516) and NFAT (5861) from cell signaling (Danvers, MA).
Jeong J., Lee J., Talaia G., Kim W., Song J., Hong J., Yoo K., Gonzalez D., Athonvarangkul D., Shin J., Dann P., Haberman A., Kim L.K., Ferguson S., Choi J, & Wysolmerski J. (2023). Intracellular Calcium links Milk Stasis to Lysosome Dependent Cell Death by Activating a TGFβ3/TFEB/STAT3 Pathway Early during Mammary Gland Involution. Research Square.
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