Western Blot:Article Title: Ring finger protein 213 regulates B-cell receptor signaling, metabolism, and development in B lymphocytes
Article Snippet: Subsequently, cell lysis was carried out with RIPA lysis buffer (Beyotime, P0013B). .. The cell lysates were examined via SDS-PAGE, and immunoblotting was performed with the following antibodies: anti-pSYK (Cell Signaling Technology, CST, 2710S), anti-SYK (CST, 13198S), anti-pCD19 (CST, 3571S), anti-CD19 (CST, 90176S), anti-β-actin (Proteintech, 66009-1-Ig), anti-pNFκB pP65 (CST, 3033S), anti-NFκB P65 (CST, 4764S), anti-pSTAT1 (CST, 9167S), anti-STAT1 (CST, 14994S), anti-pSTAT3 (CST, 9145S), anti-STAT3 (CST, 9139S), anti-pSTAT5 (CST, 4322S), anti-STAT5 (CST, 94205S), anti-pPI3K pP85/pP55 (CST, 4228 L), anti-PI3K P85 (CST, 4292S), anti-pAKT (CST, 4060 L), anti-AKT (CST, 9272S), anti-pmTOR (CST, 5536S), anti-mTOR (CST, 2983S), anti-pS6 (CST, 4858 L), anti-S6 (CST, 2217S), anti-pFOXO1 (CST, 9461S), anti-FOXO1 (CST, 2880S), anti-CTCF (ABclonal, A19588), anti-SPIB (CST, 14337S), anti-ETS1 (ZenBio, R382074), anti-GABPA (HuaBio, HA500187 ), anti-FLI1 (HuaBio, HA500149 ), anti-PU.1 (ABclonal, A20461), anti-GAPDH (Proteintech, 60004-1-Ig), anti-PIK3C3 (ABclonal, A12295), anti-PTEN (CST, 9188S), anti-pSHIP-1 (CST, 3941S), SHIP-1 (ABclonal, A3571), anti-NOTCH2 (ABclonal, A24877), anti-pHSL (CST, 4126S), anti-HSL (CST, 4107S), anti-ATGL (CST, 2138S), anti-LC3 I/II (ABclonal, A5618), IκBα (ABclonal, A19714), pIκBα (SelleckChem, F2237), pIKKα/β (CST, 2697S), pP38 (SelleckChem, F0159), HSP90 (ABclonal, A5027), pJNK1/2 (ABclonal, AP0473), pERK1/2 (ABclonal, AP0472). ..
Article Title: Ring finger protein 213 regulates B-cell receptor signaling, metabolism, and development in B lymphocytes.
Article Snippet: Subsequently, cell lysis was carried out with RIPA lysis buffer (Beyotime, P0013B). .. The cell lysates were examined via SDS-PAGE, and immunoblotting was performed with the following antibodies: anti-pSYK (Cell Signaling Technology, CST, 2710S), antiSYK (CST, 13198S), anti-pCD19 (CST, 3571S), anti-CD19 (CST, 90176S), anti-β-actin (Proteintech, 66009-1-Ig), anti-pNFκB pP65 (CST, 3033S), anti-NFκB P65 (CST, 4764S), anti-pSTAT1 (CST, 9167S), anti-STAT1 (CST, 14994S), anti-pSTAT3 (CST, 9145S), anti-STAT3 (CST, 9139S), anti-pSTAT5 (CST, 4322S), anti-STAT5 (CST, 94205S), anti-pPI3K pP85/pP55 (CST, 4228 L), anti-PI3K P85 (CST, 4292S), anti-pAKT (CST, 4060 L), anti-AKT (CST, 9272S), anti-pmTOR (CST, 5536S), anti-mTOR (CST, 2983S), anti-pS6 (CST, 4858 L), anti-S6 (CST, 2217S), anti-pFOXO1 (CST, 9461S), anti-FOXO1 (CST, 2880S), anti-CTCF (ABclonal, A19588), anti-SPIB (CST, 14337S), anti-ETS1 (ZenBio, R382074), anti-GABPA (HuaBio, HA500187), anti-FLI1 (HuaBio, HA500149), anti-PU.1 (ABclonal, A20461), anti-GAPDH (Proteintech, 60004-1-Ig), anti-PIK3C3 (ABclonal, A12295), antiPTEN (CST, 9188S), anti-pSHIP-1 (CST, 3941S), SHIP-1 (ABclonal, A3571), anti-NOTCH2 (ABclonal, A24877), anti-pHSL (CST, 4126S), anti-HSL (CST, 4107S), anti-ATGL (CST, 2138S), anti-LC3 I/II (ABclonal, A5618), IκBα (ABclonal, A19714), pIκBα (SelleckChem, F2237), pIKKα/β (CST, 2697S), pP38 (SelleckChem, F0159), HSP90 (ABclonal, A5027), pJNK1/2 (ABclonal, AP0473), pERK1/2 (ABclonal, AP0472). ..
Electrophoresis:Article Title: SARS-CoV-2 infection causes immunodeficiency in recovered patients by downregulating CD19 expression in B cells via enhancing B-cell metabolism
Article Snippet: .. Cell lysates were used for electrophoresis in SDS–polyacrylamide gel, electrotransferred onto a nitrocellulose membrane, and then probed with the following specific antibodies: anti-pCD19 (Cat# 3571S, Cell Signaling Technology, USA), anti-CD19 (Cat# 90176, Cell Signaling Technology, USA), anti-pBtk (Cat# ab52192, Abcam, USA), anti-Btk (Cat# 8547S, Cell Signaling Technology, USA), anti-pPI3K (Cat# 4228S, Cell Signaling Technology, USA), anti-pAkt (Cat# 4060L, Cell Signaling Technology, USA), anti-pFoxO1 (Cat# 9461S, Cell Signaling Technology, USA), anti-pmTOR (Cat# 5536S, Cell Signaling Technology, USA), and anti-pS6 (Cat# 4856S, Cell Signaling Technology, USA). .. After incubated relative secondary antibodies, immunoreactive bands were presented and captured with the ChemiDoc XRS + imaging systems (Bio-Rad). β-actin or GAPDH was used as the loading control.
Membrane:Article Title: SARS-CoV-2 infection causes immunodeficiency in recovered patients by downregulating CD19 expression in B cells via enhancing B-cell metabolism
Article Snippet: .. Cell lysates were used for electrophoresis in SDS–polyacrylamide gel, electrotransferred onto a nitrocellulose membrane, and then probed with the following specific antibodies: anti-pCD19 (Cat# 3571S, Cell Signaling Technology, USA), anti-CD19 (Cat# 90176, Cell Signaling Technology, USA), anti-pBtk (Cat# ab52192, Abcam, USA), anti-Btk (Cat# 8547S, Cell Signaling Technology, USA), anti-pPI3K (Cat# 4228S, Cell Signaling Technology, USA), anti-pAkt (Cat# 4060L, Cell Signaling Technology, USA), anti-pFoxO1 (Cat# 9461S, Cell Signaling Technology, USA), anti-pmTOR (Cat# 5536S, Cell Signaling Technology, USA), and anti-pS6 (Cat# 4856S, Cell Signaling Technology, USA). .. After incubated relative secondary antibodies, immunoreactive bands were presented and captured with the ChemiDoc XRS + imaging systems (Bio-Rad). β-actin or GAPDH was used as the loading control.
Article Title: HMGB1 couples LEF1 to regulate B cell immunity.
Article Snippet: Images were captured using the Nikon TIRFm system (Nikon Eclipse Ti-PFS) with 405, 488, 546, and 647 nm lasers, and colocalization and MFI were determined with the NIS-Elements AR 3.2 software. .. SDS-PAGE loading buffer (5×) (BL502A; Biosharp) was added to the cell lysates, and proteins were separated using SDS-PAGE gel and transferred to a nitrocellulose membrane for detection using the following antibodies: anti-pCD19 (3571S; CST), anti-CD19 (90176S; CST), anti-pY, anti-pBTK (5082S; CST), anti-BTK (8547S; CST), anti-pSHIP-1 (3941S; CST), anti-SHIP-1 (2728S; CST), anti-pPI3K (4228S; CST), anti-PI3K (4292S; CST), anti-pAKT (4060S; CST), anti-AKT (9272S; CST), anti-pFOXO1 (9461S; CST), anti-FOXO1 (2880S; CST), anti-pS6 (4856S; CST), anti-S6 (2217S; CST), anti-pmTOR (5536S; CST), anti-mTOR (5536S; CST), anti-pWASP (ab59278; Abcam), anti-WASP (sc13139; Santa Cruz Biotechnology), anti-pEzrin (3726S; CST), anti-WIP (sc271113; Santa Cruz Biotechnology), antiDOCK8 (sc292124; Santa Cruz Biotechnology), anti-pMST1 (3681S; CST), anti-MST1 (PA5-22015; 1 7 R E S E A R C H A R T I C L E JCI Insight 2025;10(18):e187002 https://doi.org/10.1172/jci.insight.187002 Figure 8. .. Splenocytes were incubated with or without biotin-conjugated F(ab′)2 Ig (M+G) (115- 066-068; Jackson ImmunoResearch) for 30 minutes and streptavidin (16000114; Jackson ImmunoResearch) for 10 minutes on ice, then stimulated for 5, 10, and 30 minutes at 37°C, except for those in the resting stage.
other:Article Title: HMGB1 couples LEF1 to regulate B cell immunity
Article Snippet: Cells were then lysed using RIPA lysis buffer (Beyotime; P0013B) containing protease inhibitor cocktail (1×) (G2006; Servicebio), 10 mM NaF (G2007-1; Servicebio), and 1 mM Na 3 VO 3 (G2007-2; Servicebio).
Staining:
SDS Page:Article Title: HMGB1 couples LEF1 to regulate B cell immunity.
Article Snippet: Images were captured using the Nikon TIRFm system (Nikon Eclipse Ti-PFS) with 405, 488, 546, and 647 nm lasers, and colocalization and MFI were determined with the NIS-Elements AR 3.2 software. .. SDS-PAGE loading buffer (5×) (BL502A; Biosharp) was added to the cell lysates, and proteins were separated using SDS-PAGE gel and transferred to a nitrocellulose membrane for detection using the following antibodies: anti-pCD19 (3571S; CST), anti-CD19 (90176S; CST), anti-pY, anti-pBTK (5082S; CST), anti-BTK (8547S; CST), anti-pSHIP-1 (3941S; CST), anti-SHIP-1 (2728S; CST), anti-pPI3K (4228S; CST), anti-PI3K (4292S; CST), anti-pAKT (4060S; CST), anti-AKT (9272S; CST), anti-pFOXO1 (9461S; CST), anti-FOXO1 (2880S; CST), anti-pS6 (4856S; CST), anti-S6 (2217S; CST), anti-pmTOR (5536S; CST), anti-mTOR (5536S; CST), anti-pWASP (ab59278; Abcam), anti-WASP (sc13139; Santa Cruz Biotechnology), anti-pEzrin (3726S; CST), anti-WIP (sc271113; Santa Cruz Biotechnology), antiDOCK8 (sc292124; Santa Cruz Biotechnology), anti-pMST1 (3681S; CST), anti-MST1 (PA5-22015; 1 7 R E S E A R C H A R T I C L E JCI Insight 2025;10(18):e187002 https://doi.org/10.1172/jci.insight.187002 Figure 8. .. Splenocytes were incubated with or without biotin-conjugated F(ab′)2 Ig (M+G) (115- 066-068; Jackson ImmunoResearch) for 30 minutes and streptavidin (16000114; Jackson ImmunoResearch) for 10 minutes on ice, then stimulated for 5, 10, and 30 minutes at 37°C, except for those in the resting stage.
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