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Mouse monoclonal anti dnak

Manufactured by Enzo Life Sciences
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Mouse monoclonal anti-DnaK is a laboratory reagent used to detect the presence of the DnaK protein. DnaK is a chaperone protein found in many organisms, including bacteria. This antibody can be used to identify and study the DnaK protein in various experimental settings.

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Lab products found in correlation

Mouse monoclonal anti flag epitope clone m2, by Merck Group (1 mentions) Anti rabbit igg conjugated to horseradish peroxidase, by Bio-Rad (1 mentions) 0.45 μm nitrocellulose membrane, by Bio-Rad (1 mentions) Azure c600 imaging system, by Azure Biosystems (1 mentions)

Close spelling variants of this product

Mouse monoclonal anti-DnaK (8E2/2, Mouse anti‐DnaK Anti-DnaK

2 protocols using mouse monoclonal anti dnak

1

Western Blot Antibody Dilutions

Cited 2 times
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Primary antibodies and their working dilutions used for western blotting included: mouse monoclonal anti-FLAG epitope (clone M2, Sigma) 1:5,000; rabbit polyclonal anti-IgaA (Cano et al., 2002 (link)) 1:10,000; rabbit polyclonal anti-OmpA (gift from H. Schwarz, University of Tübingen, Germany) 1:50,000; mouse monoclonal anti-DnaK (Enzo) 1:10,000; mouse monoclonal anti-β-lactamase (gift from L.A. Fernández, CNB-CSIC, Madrid, Spain) 1:2,000; rabbit polyclonal anti-PBP3 (Castanheira et al., 2017 (link)) 1:1,000; and, rabbit polyclonal anti-PBP2 (Castanheira et al., 2020 (link)) 1:1,000. Goat polyclonal anti-mouse or anti-rabbit IgG conjugated to horseradish peroxidase (Bio-Rad) were used as secondary antibodies at a 1:10,000 dilution. SDS-PAGE and western blotting were performed as described (Nunez-Hernandez et al., 2014 (link)).
Cestero J.J., Castanheira S., Pucciarelli M.G, & García-del Portillo F. (2021). A Novel Salmonella Periplasmic Protein Controlling Cell Wall Homeostasis and Virulence. Frontiers in Microbiology, 12, 633701.
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2

SDS-PAGE and Western Blot Analysis

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Bacterial lysates were solubilized in boiling 1.5X Laemmli sample buffer
and stored at −20°C until ready for analysis. Proteins were
separated by 10% SDS-PAGE (v/v bis-acrylamide gels) and transferred to
0.45μm nitrocellulose membranes (Bio-Rad Laboratories). Membranes were
processed for immunodetection with the SNAP i.d. system (EMD Millipore).
Membranes were blocked in Tris-buffered saline containing 0.5% (w/v) milk powder
and 0.1% (v/v) Tween 20 (TBS-T), incubated with primary antibodies, mouse
monoclonal anti-SipA 1A9 (1:50, Finn et al.
2017) and mouse monoclonal anti-DnaK (1:3500; clone 8E2/2; Enzo),
washed three times with TBS-T, then incubated with horseradish peroxidase-linked
anti-mouse secondary antibodies (1:3500; Cell Signaling Technology), followed by
another three washes with TBS-T. Blots were developed using the SuperSignal
Femto Substrate Kit (Thermo Fisher Scientific) according to the
manufacturer’s instruction. Images were captured with an Azure c600
Imaging System (Azure Biosystems) and analyzed for densitometry with ImageJ
software (W.S. Rasband, NIH, version 1.51).
Chong A., Starr T., Finn C.E, & Steele-Mortimer O. (2019). A role for the Salmonella Type III Secretion System 1 in bacterial adaptation to the cytosol of epithelial cells. Molecular microbiology, 112(4), 1270-1283.
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