Cytospin slides were fixed in paraformaldehyde 4% (Sigma-Aldrich). After 3 lavages in TBS, cells were incubated 10 min in TBS-0.3% Triton X-100, were washed 3 times in TBS, blocked in TBS-1% BSA-10% SVF during 45 min and incubated with primary anti-BAFF (Abcam, ab16081) or control isotype antibodies over night at 4°C. After 3 lavages in TBS, cells were incubated 1 h at room temperature with anti-rat IgM conjugated to Alexa 488. Cytospins were counterstained using 4′,6-diamidino-2-phenylindole (DAPI) for 10 min, rinsed and coverslip were mounted with Mowiol (Sigma-Aldrich). For fluorescent intensity quantification, the gray value was calculated in each cell selected on untreated pictures. Cells were observed using a Nikon eclipse 80i microscope and images were treated using ImageJ software.
Ab16081
Manufactured by Abcam
Sourced in United States
Ab16081 is a rabbit polyclonal antibody that specifically recognizes the human Axl protein. Axl is a receptor tyrosine kinase involved in cell survival, proliferation, and migration. This antibody can be used for various applications such as Western blotting, immunoprecipitation, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Mowiol, by Merck Group (1 mentions)
Paraformaldehyde 4, by Merck Group (1 mentions)
Eclipse 80i microscope, by Nikon (1 mentions)
Kgp902, by Keygen Biotech (1 mentions)
Ab8227, by Abcam (1 mentions)
Ab85803, by Abcam (1 mentions)
Immobilon p, by Merck Group (1 mentions)
Bradford protein assay, by Bio-Rad (1 mentions)
Anti β actin sc 47778, by Santa Cruz Biotechnology (1 mentions)
3 protocols using ab16081
1
Immunofluorescence Detection of BAFF
2
Western Blot Analysis of BAFF Protein
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Total cells were homogenized, and total proteins were extracted using a protein extraction kit (KGP902, KeyGen Biotech). Equal amounts of protein were separated via 15% SDS‐PAGE and transferred to polyvinylidene fluoride membranes. Subsequently, the membranes were blocked with 5% non‐fat milk for 2 h and incubated with anti‐BAFF (ab16081, Abcam) and anti‐β‐actin (ab8227, Abcam) antibodies at 4°C overnight. The membranes were washed thrice with TBST buffer and incubated with horseradish peroxidase‐conjugated secondary antibodies. Detection was visualized using an enhanced chemiluminescence system.
3
Western Blot Analysis of Immune Signaling
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Tissues were lysed in Pro-Prep solution (Intron, Seongnam, Korea). Total protein was measured using the Bradford protein assay (BioRad Laboratories). Proteins were separated by 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto polyvinylidene difluoride membranes (Immobilon-P; Millipore, Burlington, MA, USA). Membranes were blocked with the Universal WB blocking buffer (JUBIOTECH, Deajeon, Korea) and incubated with the appropriate primary antibodies. Blots were then incubated with the appropriate peroxidase-conjugated secondary antibodies. The membranes were developed using a chemiluminescent reagent (enhanced chemiluminescent; GE Healthcare, Chicago, IL, USA) and subsequently exposed to X-ray film to visualize the signal. The following primary antibodies were used in western blotting analysis: IFN-α (F-7; Santa Cruz, Dallas, TX, USA), IFN-β (ab85803; Abcam, Cambridge, MA, USA), BAFF (ab16081; Abcam), p65 (F-6; Santa Cruz), and anti-β actin (sc-47778; Santa Cruz).
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