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Anti pgrmc1

Manufactured by Cell Signaling Technology
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Anti-PGRMC1 is a laboratory reagent used for the detection and analysis of the PGRMC1 protein. PGRMC1 is a membrane-associated progesterone receptor involved in cellular signaling processes. This antibody product can be used in various research applications, such as Western blotting and immunohistochemistry, to study the expression and localization of PGRMC1 in biological samples.

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Lab products found in correlation

Anti β actin, by Cell Signaling Technology (2 mentions) Anti gapdh, by Merck Group (1 mentions) Protease and phosphatase inhibitor, by Cell Signaling Technology (1 mentions) Roti fluoro, by Carl Roth (1 mentions) Anti itgb1, by Proteintech (1 mentions) Biometra fastblot, by Analytik Jena (1 mentions) Alexa fluor 488, by Cell Signaling Technology (1 mentions) Immobilon p, by Merck Group (1 mentions) Chemostar imaging system, by Intas (1 mentions) Halt protease inhibitor cocktail edta free 100x, by Thermo Fisher Scientific (1 mentions) Ecl western blotting detection system, by PerkinElmer (1 mentions) Anti egfr, by Cell Signaling Technology (1 mentions) Axioplan 2, by Zeiss (1 mentions) Axiovision software, by Zeiss (1 mentions) Anti vimentin, by Merck Group (1 mentions) Power block, by BioGenex (1 mentions)

Close spelling variants of this product

PGRMC1

4 protocols using anti pgrmc1

1

Immunoblotting Protocol for PGRMC1 Detection in Retina and Eyecup

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To detect PGRMC1 levels in retina and eyecup, tissue lysates were subjected to immunoblotting as described (13 ). Antibodies included anti-PGRMC1 (1:200, Cell Signaling Technology, Denvers, MA, USA), anti-σR1 (1:500) (50 (link)), and anti-GAPDH (1:2000, Millipore, Billerica, MA, USA) used as a loading control. Tissue lysates were prepared by sonication in cell lysis buffer with protease inhibitors. Protein samples were fractionated on SDS-PAGE gels and transferred to nitrocellulose membranes. Membranes were blocked with 5% non-fat dry milk and exposed to primary antibody at 4°C overnight followed by appropriate secondary conjugated antibody to horseradish peroxidase (1:5000, Santa Cruz Corp., Santa Cruz, CA) at room temperature for 1 h, and developed by Chemiluminescence SuperSignal Western system. Protein levels were quantified by densitometry and expressed as a ratio to GAPDH.
Shanmugam A.K., Mysona B.A., Wang J., Zhao J., Tawfik A., Sanders A., Markand S., Zorrilla E., Ganapathy V., Bollinger K.E, & Smith S.B. (2015). Progesterone receptor membrane component 1 (PGRMC1) expression in murine retina. Current eye research, 41(8), 1105-1112.
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2

Protein Expression Analysis in GBM Cells

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GBM cells were subjected to lysis using a buffer containing Triton X-100 along with protease and phosphatase inhibitors (Cell Signaling Technology). The lysates were cleared by cellular debris through centrifugation and then mixed with a loading buffer containing 4% glycerin, 0.8% SDS, 1.6% beta-mercaptoethanol, and 0.04% bromophenol blue (all from Carl Roth). The proteins were electrophoretically separated by SDS-PAGE and then transferred to Immobilon-P (Merck Millipore, Darmstadt, Germany) or Roti®-Fluoro (Carl Roth) PVDF membranes using a semidry blotting device (Biometra FastblotTM, Analytik Jena AG, Jena, Germany). The membranes were incubated overnight at 4 °C with the following primary antibodies: anti-PGRMC1, anti-TCF 1/7, anti-beta-Actin (all from Cell Signaling Technology), or anti-ITGB1 (Proteintech, Manchester, UK). Subsequent secondary reactions were carried out for 1 h at room temperature with HRP-, AlexaFluor®488-, or AlexaFluor®647-coupled antibodies (all from Cell Signaling Technology). All antibodies were diluted in SignalBoost™ Immunoreaction Enhancer (Merck Millipore) according to the manufacturer’s recommendation. The ChemoStar imaging system (Intas Science Imaging, Göttingen, Germany) was used for the detection and acquisition of signals and the intensity of the bands was quantified with the ImageJ 1.48v software.
Dumitru C.A., Schröder H., Schäfer F.T., Aust J.F., Kreße N., Siebert C.L., Stein K.P., Haghikia A., Wilkens L., Mawrin C, & Sandalcioglu I.E. (2023). Progesterone Receptor Membrane Component 1 (PGRMC1) Modulates Tumour Progression, the Immune Microenvironment and the Response to Therapy in Glioblastoma. Cells, 12(20), 2498.
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3

Western Blot Analysis of Protein Targets

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Total proteins were extracted from MCF10A using M-PER containing Halt™ Protease Inhibitor Cocktail, EDTA-Free (100X) (Thermo Fisher Scientific, Inc.). Western blot was performed as previously described [22] (link). All antibodies used in this study were purchased from Cell Signaling Technology, Inc., including anti-PGRMC1 (#13,856, l:1000), anti-EGFR (#2085, l:1000), anti-mechanistic target of rapamycin (mTOR) (#2983, l:1000), anti-phosphatase and tensin homolog (PTEN) (#9188, l:1000), and anti-β-actin (#3700, l:1000). β-actin was used as a loading control. Proteins were visualized using an ECL Western blotting detection system (PerkinElmer, Inc.). Densitometry values were measured in terms of pixel intensity by ImageJ.
Cai G., Wang Y., Houda T., Yang C., Wang L., Gu M., Mueck A., Croteau S., Ruan X, & Hardy P. (2021). MicroRNA-181a suppresses norethisterone-promoted tumorigenesis of breast epithelial MCF10A cells through the PGRMC1/EGFR–PI3K/Akt/mTOR signaling pathway. Translational Oncology, 14(6), 101068.
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4

Immunofluorescence Analysis of Retinal Cells

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Retinal cryosections were fixed in 4% paraformaldehyde for 10 min, washed three times with 0.1% Triton X-100 for 5 min. Slides were blocked (PowerBlock; BioGenex, San Ramon, CA) for 1 h at room temperature, incubated with primary antibodies overnight at 4 °C followed by secondary antibodies for 1 h at 25°C. Sections were examined for immunofluorescence using an Axioplan-2 fluorescent microscope (Carl Zeiss, Göttingen, Germany) equipped with a high-resolution microscope camera. Images were captured and processed using Zeiss Axiovision software (version 4.7; Carl Zeiss). Antibodies included anti-PGRMC1 (1:200, Cell Signaling Technology, Denvers, MA, USA), anti-vimentin (1:200, Millipore, Billerica, MA, USA) to label retinal Müller cells, and anti-Brn3a, a marker for neurons (1:100; Santa Cruz Corp., Santa Cruz, CA). Secondary antibodies (Alexafluor 488 and Alexafluor 555) were obtained from Invitrogen-Life Sciences (Grand Island, NY).
Shanmugam A.K., Mysona B.A., Wang J., Zhao J., Tawfik A., Sanders A., Markand S., Zorrilla E., Ganapathy V., Bollinger K.E, & Smith S.B. (2015). Progesterone receptor membrane component 1 (PGRMC1) expression in murine retina. Current eye research, 41(8), 1105-1112.
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