Cell surface markers were stained in 1×PBS containing 2% FBS with indicated antibodies. Anti-CD4 (RM4-5), anti-CD8 (53-6.7), anti-CD25 (PC61), anti-CD44 (IM7), anti-CD62L (MEL-14), anti-CD45.1 (A20), and anti-CD45.2 (104) were purchased from eBioscience. Intracellular staining was performed using the Foxp3/Transcription Factor Staining Kit (eBioscience) with indicated antibodies. For intracellular cytokine staining, cells were stimulated for 4 h with phorbol 12-myristate 13-acetate (50 ng/mL; Sigma-Aldrich) and ionomycin (1 μg/mL; Sigma-Aldrich), and then treated for another 1 h with Golgi-Plug (BD Biosciences). Anti–IFN-γ (XMG1.2), anti-IL17A (eBio17B7), and anti–IL-4 (eBio 11B11) were purchased from eBioscience, and anti–cleaved caspase-3(Asp175) (D3E9), anti–phospho-H2A.x(Ser139) (20E3), and anti-LC3b (D11) were purchased from Cell Signaling Technology. All data collection was performed with a FACSAria II cell sorter (BD Biosciences), and data analysis was done with FlowJo software (Tree Star).
Anti lc3b d11
Manufactured by Cell Signaling Technology
Anti-LC3b (D11) is a rabbit monoclonal antibody that recognizes the light chain 3B (LC3B) protein. LC3B is a marker for autophagosome formation and a key protein involved in the autophagy process. This antibody can be used to detect LC3B expression in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Foxp3 transcription factor staining kit, by Thermo Fisher Scientific (1 mentions)
Anti cd45.2 104, by Thermo Fisher Scientific (1 mentions)
Anti ifnγ xmg1.2, by Thermo Fisher Scientific (1 mentions)
Anti il 17a ebio17b7, by Thermo Fisher Scientific (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
Facsaria 2 cell sorter, by BD (1 mentions)
Golgiplug, by BD (1 mentions)
Axioimager microscope, by Zeiss (1 mentions)
Goat anti rabbit igg, by Merck Group (1 mentions)
Ab141382, by Abcam (1 mentions)
Secondary hrp conjugated goat anti mouse igg, by Merck Group (1 mentions)
3 protocols using anti lc3b d11
1
Multiparametric Flow Cytometry Analysis
2
Quantifying Autophagy via LC3 Puncta
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Cells were fixed with 2% paraformaldehyde in PBS for 10 min at 4 °C and permeabilized with acetone for 10 min at room temperature. Blocking was performed in 5% BSA, and primary antibodies were incubated overnight at 4 °C. Anti‐LC3B(D11) was from Cell Signaling Technology (Ozyme). After incubation for 1 h with conjugated secondary antibodies, cell nuclei were counterstained with Hoechst. Cells were visualized with an Axioimager microscope (Carl Zeiss) and the axiovision ® software at a 100× magnification and processed with imagej software. Autophagy was measured by visual counting of LC3 punctua (with 50 randomly selected cells for each condition).
3
Caspase-8 Antibody Characterization in HSV-1 Infection
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Caspase-8 (human) monoclonal antibody (12F5; ALX-804-242) directed against the p18 subunit was purchased from Enzo Life Sciences. Monoclonal anti-US11 and anti-ICP8 were provided by professor Bernad Roizman. Anti-GAPDH (sc-32233), anti-HSV-1 UL42 (sc-53333) and goat anti-mouse IgG, F(ab')2-PE (sc-3798) and anti-Atg3 (sc-393660) were purchased from Santa Cruz Biotechnology. Anti-caspase 3 (#9662), anti-PARP (#9542), GAPDH (Rabbit mAb #2118) and anti-LC3B(D11) (Rabbit mAb #3868) were provided from Cell Signaling Technology. Secondary HRP-conjugated anti-mouse IgGVeriBlot for IP were from Abcam. Secondary HRP-conjugated goat anti-mouse IgG and goat anti-rabbit IgG were purchased from Millipore. The z-IETD-FMK caspase-8 inhibitor (ab141382) was purchased from Abcam.
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