Agents were purchased/obtained as follows: (1) bortezomib (Biovision, Palo Alto, CA); (2) fumonisin B1, palmitate, 4-PBA (4-phenylbutyric acid), TUDCA (tauroursodeoxycholic acid), and anti-α-tubulin, anti-HA and anti-CerS2 antibodies (Sigma-Aldrich, St Louis, MO); (3) ceramides (fatty acyl lengths C16, C18, C20, C22, and C24) and C17 ceramide (d17:1/C18:0) (Avanti Polar Lipid, Alabaster, AL); (4) anti-GRP78, anti-CHOP, anti-FAS, anti-phospho-eiF2α, anti-eiF2α, anti-PERK, and anti-SCD-1 antibodies (Cell Signaling Technology, Beverly, MA); (5) anti-SREBP-1, anti-phospho-PERK and anti-CerS6 antibodies (Santa Cruz Biotechnology, Santa Cruz, CA); (6) anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase) antibody (Millipore, Temecula, CA); (7) anti-INSIG-1 antibody (Abcam, Cambridge, MA); (8) anti-mouse-HRP (horseradish peroxidase) and anti-rabbit-HRP antibodies (Jackson Laboratory, Bar Harbor, ME).
Anti rabbit hrp antibodies
Manufactured by Jackson ImmunoResearch
Anti-rabbit HRP antibodies are secondary antibodies conjugated with horseradish peroxidase (HRP) enzyme. They are designed to detect and bind to primary antibodies raised in rabbits, allowing for the visualization and amplification of target signals in various immunoassays and immunohistochemical applications.
Automatically generated - may contain errors
Lab products found in correlation
Gel doc xr, by Bio-Rad (2 mentions)
Bortezomib, by Abcam (1 mentions)
Anti chop, by Cell Signaling Technology (1 mentions)
Anti phospho eif2α, by Cell Signaling Technology (1 mentions)
Anti scd1, by Cell Signaling Technology (1 mentions)
Fumonisin b1, by Merck Group (1 mentions)
Tudca, by Merck Group (1 mentions)
Anti grp78, by Cell Signaling Technology (1 mentions)
Anti fas, by Cell Signaling Technology (1 mentions)
Anti srebp 1, by Santa Cruz Biotechnology (1 mentions)
Anti phospho perk, by Santa Cruz Biotechnology (1 mentions)
Anti cers6 antibodies, by Santa Cruz Biotechnology (1 mentions)
Anti α tubulin, by Merck Group (1 mentions)
Anti p erk, by Cell Signaling Technology (1 mentions)
Anti gapdh, by Merck Group (1 mentions)
Anti eif2α, by Cell Signaling Technology (1 mentions)
Protease and phosphatase inhibitor cocktail, by Roche (1 mentions)
4 protocols using anti rabbit hrp antibodies
1
Ceramide Regulation of Cellular Stress
2
NFκB Signaling Pathway Analysis
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For analysis of NFκB signaling pathways, the BMDMs were sub-cultured in 12-well cell culture plates for 16 h, and stimulated with live Aspergillus spores at 25 MOI of infection for indicated times. Protein lysates were prepared using the lysis buffer (10-mM Tris–HCl, 150-mM NaCl, 1% Nonidet P-40, supplemented with protease and phosphatase inhibitor cocktails; Roche). Protein samples were denatured by boiling in sample loading buffer-containing SDS and 100-mM DTT for 5 min and separated in denaturing SDS-PAGE. Separated proteins were transferred to PVDF membranes and immunoblotted with rabbit antibodies against total IκBa, Phospho-IκBa. All antibodies were purchased from Cell Signaling followed by secondary anti-rabbit HRP antibodies (JacksonImmunoResearch Laboratories).
3
Immunoblot Analysis of Protein Depletion
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Samples were combined with 5X Laemmli sample buffer (10% SDS, 50% glycerol, 300mM Tris HCl pH 6.8, 0.05% bromophenol blue, 5% beta-mercaptoethanol) and were incubated at 95°C for 10 min. The samples were run on precast 4–15% or 7.5% SDS gels (Bio-Rad) and were transferred overnight onto a nitrocellulose membrane in transfer buffer (25mM TrisHCl, 192 mM glycine, 0.1% SDS, 20% methanol) at 4°C. Blocking was performed with 5% milk in PBS for 1 hr rocking at room temperature. Antibody incubations were performed with 5% milk in TBS-T for 1 hr rocking at room temperature. Three 5 min TBS-T washes were performed before and after secondary antibody incubations rocking at room temperature. After a final PBS wash, imaging was performed using a LI-COR Odyssey.
For immunoblot detection of the HA tag in AID-HOOK and FTS-AID after auxin-mediated depletion, secondary antibody incubations were performed with anti-rabbit HRP antibodies (Jackson ImmunoResearch) and detected with chemiluminescence (Azure) for increased sensitivity. Imaging was performed using a Bio-Rad Gel Doc XR.
For immunoblot detection of the HA tag in AID-HOOK and FTS-AID after auxin-mediated depletion, secondary antibody incubations were performed with anti-rabbit HRP antibodies (Jackson ImmunoResearch) and detected with chemiluminescence (Azure) for increased sensitivity. Imaging was performed using a Bio-Rad Gel Doc XR.
4
Western Blot Analysis of Auxin-Mediated Depletion
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Samples were combined with 5× Laemmli sample buffer (10% SDS, 50% glycerol, 300 mM Tris HCl pH 6.8, 0.05% bromophenol blue, 5% beta-mercaptoethanol) and were incubated at 95°C for 10 min. The samples were run on precast 4–15% or 7.5% SDS gels (Bio-Rad) and were transferred overnight onto a nitrocellulose membrane in transfer buffer (25 mM Tris–HCl, 192 mM glycine, 0.1% SDS, 20% methanol) at 4°C. Blocking was performed with 5% milk in PBS for 1 hr rocking at room temperature. Antibody incubations were performed with 5% milk in TBS-T for 1 hr rocking at room temperature. Three 5 min TBS-T washes were performed before and after secondary antibody incubations rocking at room temperature. After a final PBS wash, imaging was performed using a LI-COR Odyssey.
For immunoblot detection of the HA tag in AID-HOOK and FTS-AID after auxin-mediated depletion, secondary antibody incubations were performed with anti-rabbit HRP antibodies (Jackson ImmunoResearch) and detected with chemiluminescence (Azure) for increased sensitivity. Imaging was performed using a Bio-Rad Gel Doc XR.
For immunoblot detection of the HA tag in AID-HOOK and FTS-AID after auxin-mediated depletion, secondary antibody incubations were performed with anti-rabbit HRP antibodies (Jackson ImmunoResearch) and detected with chemiluminescence (Azure) for increased sensitivity. Imaging was performed using a Bio-Rad Gel Doc XR.
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