Rabbit phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP™ (#4511), rabbit VCAM-1 (#12367), rabbit phospho-ERK1/2 (#4307), rabbit phospho-JNK (#4668), rabbit phospho-cjun (#320T), rabbit β-Actin (13E5) (#4790) antibodies were from Cell Signaling Technology (Leiden, the Netherlands). Rabbit ICAM-1 (H-108) (#sc-7891) were from Santa Cruz Biotechnology (Heidelberg, Germany). Mouse GAPDH (#MAB374) as well as anti-mouse and rabbit horseradish peroxidase-conjugated secondary antibodies (#401253 and #401353) were from Merck Millipore (Darmstadt, Germany). Mouse monoclonal APC-labelled ICAM-1 (#559771), VCAM-1 (#551147), and IgG1 isotype control antibody (#555751) for flow cytometry were from BD Biosciences (Heidelberg, Germany). Recombinant human IL-1β was purchased from PeproTech. OR-1855 was purchased from Hölzel Diagnostika (Köln, Germany) and OR-1896 was purchased from Hycultec (Beutelsbach, Germany). All other chemicals were from Sigma-Aldrich (Taufkirchen, Germany).
Igg1 isotype control antibody
Manufactured by BD
Sourced in Germany, United Kingdom
The IgG1 isotype control antibody is a laboratory reagent used in immunoassay techniques. It serves as a reference for determining the specificity of target antibodies by providing a control with the same isotype but no known reactivity to the analyte of interest.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Recombinant human il 1β, by Thermo Fisher Scientific (1 mentions)
Anti mouse and rabbit horseradish peroxidase conjugated secondary antibodies, by Merck Group (1 mentions)
Anti cd81 clone js 81, by BD (1 mentions)
Goat anti rabbit igg cy5, by Abcam (1 mentions)
Pe conjugated monoclonal antibodies, by BD (1 mentions)
4 protocols using igg1 isotype control antibody
1
Rabbit Antibody Immunoblotting and Flow Cytometry
2
Investigating CD81 in Hepatocyte Infection
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Single cell suspensions of hepatocytes were incubated for 20 minutes on ice with either anti-CD81 (clone JS-81, BD Pharmingen, San Diego, CA) or IgG1 isotype control antibody, each conjugated to APC. Azide-free anti-CD81 mouse monoclonal antibody (cl.1D6) (Abcam, Cambridge, MA) or a functional grade IgG1 isotype control antibody (eBioscience, San Diego, CA) were used in CD81 blocking experiments at a final concentration of 10 μg/ml. Antibodies were either added 2 hours prior to infection and removed, added concurrently with sporozoites, or added at 6 hours pi.
3
Integrin-mediated MMP activity assay
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The broad spectrum MMP inhibitor GM6001 was from Calbiochem (Hertfordshire, UK). FITC-conjugated anti-integrin α2 (clone AK7, Abcam, Cambridge, UK), anti-integrin α3 (clone 17C6, Abcam), and IgG1 isotype control antibodies (BD Biosciences, Oxford, UK) were used in FACS assays. Anti-integrin α2β1 (clone BHA2.1) and anti-integrin α3β1 (clone M-KID2) antibodies were from Millipore (Hertfordshire, UK). Rabbit anti-MMP-1 primary antibody (Millipore) and Cy5-goat anti-rabbit IgG (Abcam) secondary antibody were used in confocal assays. Phalloidin conjugated with Alexa Fluor 594 (Thermo Fisher Scientific, Paisley, UK) was used for F-actin staining and DAPI was used as nuclear counterstain. Other reagents were purchased from Sigma-Aldrich (Dorset, UK) unless otherwise stated.
4
Quantifying PMN Activation Markers by Flow Cytometry
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Low‐affinity immunoglobulin‐Fcγ receptor IIIb (CD16b) can be found on the surface membrane of mature PMNs and can be used as a PMN identification marker. Expression of receptors on the surface of PMNs was measured by flow cytometry. The PMNs were gated according to CD16 expression and the sideward scatter profile. To study PMN activation, samples were measured for the expression of phycoerythrin (PE)‐conjugated monoclonal antibodies (mAbs) (at a final concentration of 10 μg ml−1) against CD11b and CD63 and for expression of fluorescein isothiocyanate (FITC)‐conjugated mAb against CD66b (all from BD Biosciences). Samples were incubated for 30 min on ice in the dark. Expression of cell‐surface markers was measured on the FL‐1 (Green) and FL‐2 (Red) channels and calculated as mean fluorescence intensity (MFI). Results were corrected for non‐specific binding, by subtracting the MFI values of the corresponding IgG1 isotype‐control antibodies (BD Biosciences).
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