Tissues and cells were collected and lysed with 400 μL RIPA buffer containing 1× protease inhibitors (Roche) on ice for 30 minutes. After 30 minutes, the samples were centrifugated, denaturated and separated by 12% SDS gel electrophoresis as previous.28 The rabbit anti‐IGSF23 polyclonal antibody was raised against the linear peptide sequence MQPTEAEPMEPDPTLS of the predicted human IGSF23 protein (NP_001192209), β‐actin, RANK, c‐Fos, NFATc1, p‐ERK, ERK, p38, p‐p38 antibodies were purchased from Cell Signaling Technology; mSin3A (Abcam), 5'NT, D4‐GDI and TRAP antibodies were purchased from Abcam.
D4 gdi
Manufactured by Abcam
Sourced in United Kingdom
D4-GDI is a protein that inhibits the dissociation of GDP from the Rho family of small GTPases. It plays a role in regulating the activity of these GTPases, which are involved in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by Cell Signaling Technology (2 mentions)
P erk, by Cell Signaling Technology (1 mentions)
C fos, by Cell Signaling Technology (1 mentions)
Nfatc1, by Cell Signaling Technology (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Lactate dehydrogenase, by Abcam (1 mentions)
Bca assay, by Merck Group (1 mentions)
Serpinb2, by Abcam (1 mentions)
Cathepsin k, by Abcam (1 mentions)
Mmp 9, by Cell Signaling Technology (1 mentions)
Rankl, by Santa Cruz Biotechnology (1 mentions)
2 protocols using d4 gdi
1
Protein Expression Analysis of IGSF23
2
Protein Expression Analysis of Osteoclastogenesis
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Whole cell proteins were extracted via RIPA lysis buffer (P0013B, Beyotime Co., Shanghai, China), quantified by BCA assay (Sigma-Aldrich Co), separated on NuPAGE 10%–12% polyacrylamide gels, transferred to PVDF membranes (Millipore, Billercia, MA) and blocked in 5% BSA in TBST. Incubation was performed with antibodies against Cathepsin K (Abcam, Cambridge, England, 1:1000), TRAP (Abcam:1000), MMP-9 (Cell signalling, 1:1000), Calcitonin receptor (Abcam, 1:800), SerpinB2 (Abcam, 1:800), Peroxiredoxin 6 (Abcam, 1:1000), SOD2 (Abcam, 1:2000), D4 GDI (Abcam, 1:500), Lactate Dehydrogenase (Abcam, 1:3000), OPG (Santa Cruz Biotechnology, 1:1000), RANKL (Santa Cruz Biotechnology, 1:1000), β-actin (Cell-signaling, 1:2000). After washing with TBST, blots were incubated with goat anti-rabbit, goat anti-mouse or rabbit anti-goat horseradish peroxidase-conjugated secondary antibodies previous to visualization with enhanced chemi-luminescence ECL kits (Sigma). Relative band intensities in scanned images were analyzed with Image J software.
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