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Ab90867

Manufactured by Abcam
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Sourced in United States

Ab90867 is a laboratory equipment product. It serves as a core function for specific laboratory applications. No further details are available.

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Lab products found in correlation

Ab3328, by Abcam (1 mentions) Ab3329, by Abcam (1 mentions) Stat6, by Cell Signaling Technology (1 mentions) P stat6, by Cell Signaling Technology (1 mentions) Hrp conjugated anti β actin, by Merck Group (1 mentions) Polyvinylidene difluoride membrane, by Bio-Rad (1 mentions) Ab77735, by Abcam (1 mentions) Ab155091, by Abcam (1 mentions) Ab183727, by Abcam (1 mentions) Ab7291, by Abcam (1 mentions) Ab1791, by Abcam (1 mentions) Mini protean transfer system, by Bio-Rad (1 mentions)

2 protocols using ab90867

1

Alveolar Macrophage Protein Isolation

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For protein isolation, alveolar macrophages were washed twice with ice cold PBS and lysed with RIPA buffer (50 mM Tris·HCl, pH 7.5, 150 mM NaCl, 1% Nonidet P-40, 0.5% sodium deoxycholate and 0.1% SDS). Samples were centrifuged to remove cell debris and protein concentrations were determined using standard Bradford assays. In all, 10 μg of protein lysates were separated on 10% SDS-PAGE and blotted onto polyvinylidene difluoride membranes (162–0177, Bio-Rad, Hercules, CA, USA). The following antibodies were used: LMP2 (ab3328, Abcam, Cambridge, UK), LMP7 (ab3329, Abcam), 20S α1+2+3+5+6+7 (hereafter called α1–7; clone MCP231, ab2267, Abcam), Psmb5 (detecting β5, ab90867, Abcam), AKT (4685, Cell Signaling, Beverly, MA, USA), p-AKT (4060, pSer473, Cell Signaling), STAT6 (9362, Cell Signaling), p-STAT6 (9361, pTyr641, Cell Signaling), IRF4 (M17, Santa Cruz, Dallas, TX, USA), Arginase 1 (H-52, Santa Cruz), iNOS (610331, BD Transduction Laboratories, Franklin Lakes, NJ, USA), IL4R (ab157162, Abcam), HRP-conjugated anti-β-actin (Sigma-Aldrich), HRP-conjugated anti-rabbit (Abcam) and anti-goat antibodies (Santa Cruz).
Chen S., Kammerl I.E., Vosyka O., Baumann T., Yu Y., Wu Y., Irmler M., Overkleeft H.S., Beckers J., Eickelberg O., Meiners S, & Stoeger T. (2016). Immunoproteasome dysfunction augments alternative polarization of alveolar macrophages. Cell Death and Differentiation, 23(6), 1026-1037.
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2

Proteasome Subunit Quantification by Western Blot

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Proteins were transferred to PVDF membranes using a Bio-Rad mini-protean transfer system containing a buffer of 25 mM Tris base, 192 mM glycine, 0.1% SDS. Proteins were transferred for 6h at 40V at 4°C. Membranes with transferred proteins were blocked with a 1:1 mixture of Li-COR blocking buffer and PBS. Membranes were then incubated in blocking solution for 1 h and primary Abs (1:1000) applied for 16 h at 4°C. Membranes were washed and incubated with secondary Abs (IR Dye® 800CW goat anti-mouse IgG (H+L) or IR Dye 680CW donkey anti-rabbit IgG (H+L), and imaged using an Odyssey Li-Cor platform. Antibodies used were rabbit anti-PSMB5 polyclonal (Abcam, ab90867), rabbit monoclonal to anti-PSME1 (EPR10967, Abcam, ab155091), rabbit monoclonal to anti-PSME2 (EPR14931, Abcam, ab183727), rabbit anti-LMP2 polyclonal (PSMB9, Abcam, ab42987), rabbit anti-PSMB10 polyclonal (Abcam, ab77735), rabbit anti-LMP7 monoclonal (EPR14482, PSMB8, Abcam, ab180606), rabbit anti-PSMA6 polyclonal (Cell Signaling 2459), rabbit anti-PSMB1 polyclonal (Abcam, ab196623), rabbit anti-PSMB2 polyclonal (Abcam, ab140426), mouse anti-Rpn5 monoclonal (PSMD12, clone H3, Santa Cruz Biotechnology, sc-398279), mouse anti-α-tubulin monoclonal (DM1a, Abcam, ab7291), and rabbit anti-histone 3 polyclonal (Abcam, ab1791).
Woodle E.S., Tremblay S., Brailey P., Girnita A., Alloway R.R., Aronow B., Dasgupta N., Ebstein F., Kloetzel P.M., Lee M.J., Kim K.B., Singh H, & Driscoll J.J. (2019). Proteasomal adaptations underlying carfilzomib-resistance in human bone marrow plasma cells. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 20(2), 399-410.
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