The growth complementation assays were performed as previously described74 (link). In brief, E. coli LB2003, a strain lacking all endogenous K+ uptake systems, was transformed with plasmids encoding His-tagged KdpFABC variants. LB2003 transformed with empty vector pBAD18 or plasmid pBXC3H-KdpFABD307NC, encoding for an inactive variant, served as negative controls. Growth was monitored for 24 h at different K+ concentrations (1–115 mM, referred to as K1–K115). At K10 and below, the strain only grows sufficiently if the produced protein complements the lacking K+ transport systems. Protein production was confirmed by SDS-PAGE and subsequent western blotting analysis of a K30 sample after 24 h using an anti-His antibody from mouse (dilution 1:3000, Sigma–Aldrich, cat.no. H1029) and secondary anti-mouse IgG-HRP antibody produced in goat (dilution 1:20,000, Sigma–Aldrich, cat.no. A2554).
Anti mouse igg hrp antibody
Manufactured by Merck Group
Sourced in United States
The Anti-mouse IgG-HRP antibody is a secondary antibody labeled with horseradish peroxidase (HRP) that binds to mouse immunoglobulin G (IgG) molecules. This antibody is commonly used in various immunoassay techniques, such as Western blotting and ELISA, to detect and quantify the presence of mouse proteins or antigens.
Automatically generated - may contain errors
Lab products found in correlation
Anti his antibody from mouse, by Merck Group (2 mentions)
Anti β actin, by Merck Group (1 mentions)
Foetal bovine serum (fbs), by Merck Group (1 mentions)
Complete mini edta free protease inhibitor cocktail tablet, by Roche (1 mentions)
Rpmi media, by Merck Group (1 mentions)
Anti rabbit igg hrp, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Trypsin, by Promega (1 mentions)
4 protocols using anti mouse igg hrp antibody
1
K+ Uptake System Complementation Assay
2
Investigating IL-13 and IL-33 Signaling Pathways
3
Complementation Assay for K+ Transport
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The growth complementation assays were performed as previously described 66 .
In brief, E. coli LB2003, a strain lacking all endogenous K + uptake systems, was transformed with plasmids encoding His-tagged KdpFABC variants. LB2003 transformed with empty vector pBAD18 or plasmid pBXC3H-KdpFABD307NC, encoding for an inactive variant, served as negative controls. Growth was monitored for 24 h at different K + concentrations (1-115 mM, referred to as K1-K115). At K10 and below, the strain only grows sufficiently if the produced protein complements the lacking K + transport systems. Protein production was confirmed by SDS-PAGE and subsequent Western blotting analysis of a K30 sample after 24 h using an anti-His antibody from mouse (dilution 1:3000, Sigma-Aldrich, cat.no.
H1029) and secondary anti-mouse IgG-HRP antibody produced in goat (dilution 1:20,000, Sigma-Aldrich, cat.no. A2554).
In brief, E. coli LB2003, a strain lacking all endogenous K + uptake systems, was transformed with plasmids encoding His-tagged KdpFABC variants. LB2003 transformed with empty vector pBAD18 or plasmid pBXC3H-KdpFABD307NC, encoding for an inactive variant, served as negative controls. Growth was monitored for 24 h at different K + concentrations (1-115 mM, referred to as K1-K115). At K10 and below, the strain only grows sufficiently if the produced protein complements the lacking K + transport systems. Protein production was confirmed by SDS-PAGE and subsequent Western blotting analysis of a K30 sample after 24 h using an anti-His antibody from mouse (dilution 1:3000, Sigma-Aldrich, cat.no.
H1029) and secondary anti-mouse IgG-HRP antibody produced in goat (dilution 1:20,000, Sigma-Aldrich, cat.no. A2554).
4
Affinity-based Protein Purification and Western Blot Analysis
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Cibacron F3GA agarose, DEAE-cellulose gel beads, anti-human AGP-1 antibody, anti-mouse-IgG-HRP antibody, commercial AGP-1, Foetal Bovine Serum (FBS), LPS and RPMI media were procured from Sigma Chemicals Co., St. Louis, MO, USA. Phospho-specific antibodies to p-38, JNK and ERK, β-actin and anti-rabbit IgG-HRP were obtained from Cell Signaling Technology, Danvers, MA, USA. Complete Mini EDTA-free protease inhibitor cocktail tablets were from Roche Diagnostics, Mannheim, Germany. PVDF membrane was from BioRad Laboratories, Hercules, California, USA and Thioglycollate media was obtained from Sisco Research Laboratories, Mumbai, India. Trypsin was bought from Promega, Madison, WI, USA.
Calcein 2 AM and molecular weight markers were procured from Invitrogen, Carlsbad, CA, USA.
Calcein 2 AM and molecular weight markers were procured from Invitrogen, Carlsbad, CA, USA.
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