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Ab66506

Manufactured by Abcam
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Ab66506 is a primary antibody product from Abcam. It is designed for use in various laboratory applications.

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Lab products found in correlation

Alexa 647, by Thermo Fisher Scientific (2 mentions) Tryple, by Thermo Fisher Scientific (2 mentions) Ab30477, by BD (2 mentions) Ab82270, by Abcam (2 mentions) Ab26405, by Abcam (2 mentions) Pvdf membrane, by Merck Group (1 mentions) Protease inhibitor mixture tablet, by Roche (1 mentions)

3 protocols using ab66506

1

Macrophage Protein Expression Analysis

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Acquired macrophages (106 cells per sample) were lysed in with 50 μl buffer composing of NaCl (150 mM), Tris (50 mM, pH 7.4), 1% Nonidet P-40, 0.5% sodium deoxycholate, and complete Protease Inhibitor Mixture Tablets (Roche). Cell lysates were placed in ice for 30 min, and centrifuged at 17,000 × g, 4 °C for 30 min. After centrifugation, the supernatant was mixed with 4 × loading buffer. The samples were cooked at 95 °C for 20 min and subjected to 10% SDS/PAGE gels electrophoresis. Protein bands were transferred to PVDF membranes (EMD Millipore) at 75 V for 3 h. The membranes were blocked for 1 h at room temperature in NaCl (150 mM), Tris-HCl (10 mM, pH7.6), and 0.1% Tween 20 containing 5% nonfat milk and then probed with anti-c-Maf antibody (Santa Cruz, sc-7866) or anti-MafB (Abcam, ab66506). ECL substrate (Thermo Fisher Scientific Life Sciences) was used for protein detection.
Chen J., Du Y., He X., Huang X, & Shi Y.S. (2017). A Convenient Cas9-based Conditional Knockout Strategy for Simultaneously Targeting Multiple Genes in Mouse. Scientific Reports, 7, 517.
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2

Multicolor Flow Cytometry Analysis of Pancreatic Endocrine Cells

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Clusters at indicated stages were dissociated with TrypLE (GIBCO) with 20μg/ml DNase for 12 min at 37°C and then fixed with 4% PFA for 10 min at room temperature. Clusters were then permeabilized with 0.2 % Triton X for 10 min, blocking with 10% goat serum for 30 min and stained for various intracellular markers with antibodies, c-peptide, (1/100, abcam, ab30477), PDX-1 (1/100, BD, 562161), NKX6–1 (1/100, BD, 563338), Chromogranin A (1/100, BD, 564583), MAFA (1/100, abcam, ab26405), MAFB (1/100, abcam, ab66506), Glucagon (1/100, abcam, ab82270), Somatostatin (1/100, abcam, 108456) for analysis BD Biosciences LSRII. Data were analysed by FlowJo software. Secondary antibodies for c-peptide, Glucagon and Somatostatin were coupled to Alexa 647 (Life Technologies).
Yoshihara E., O’Connor C., Gasser E., Wei Z., Oh T.G., Tseng T.W., Wang D., Cayabyab F., Dai Y., Yu R.T., Liddle C., Atkins A.R., Downes M, & Evans R.M. (2020). Immune evasive human islet-like organoids ameliorate diabetes. Nature, 586(7830), 606-611.
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3

Multicolor Flow Cytometry Analysis of Pancreatic Endocrine Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Clusters at indicated stages were dissociated with TrypLE (GIBCO) with 20μg/ml DNase for 12 min at 37°C and then fixed with 4% PFA for 10 min at room temperature. Clusters were then permeabilized with 0.2 % Triton X for 10 min, blocking with 10% goat serum for 30 min and stained for various intracellular markers with antibodies, c-peptide, (1/100, abcam, ab30477), PDX-1 (1/100, BD, 562161), NKX6–1 (1/100, BD, 563338), Chromogranin A (1/100, BD, 564583), MAFA (1/100, abcam, ab26405), MAFB (1/100, abcam, ab66506), Glucagon (1/100, abcam, ab82270), Somatostatin (1/100, abcam, 108456) for analysis BD Biosciences LSRII. Data were analysed by FlowJo software. Secondary antibodies for c-peptide, Glucagon and Somatostatin were coupled to Alexa 647 (Life Technologies).
Yoshihara E., O’Connor C., Gasser E., Wei Z., Oh T.G., Tseng T.W., Wang D., Cayabyab F., Dai Y., Yu R.T., Liddle C., Atkins A.R., Downes M, & Evans R.M. (2020). Immune evasive human islet-like organoids ameliorate diabetes. Nature, 586(7830), 606-611.
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