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Mouse anti gapdh clone gapdh 71

Manufactured by Merck Group

Mouse anti-GAPDH (clone GAPDH-71.1) is a monoclonal antibody that recognizes the GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase) protein. GAPDH is a widely used reference protein for Western blot analysis and other immunodetection techniques.

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2 protocols using mouse anti gapdh clone gapdh 71

1

Antibody Characterization for Cellular Imaging

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The mouse monoclonal anti-GST antibody (clone 2H3-D10, diluted 1:1000) is available
from Sigma. The mouse anti-GFP antibody (clones 7.1 and 13.1, diluted 1:1000 to
1:5000) was purchased from Roche (order number 11814460001). The monoclonal anti-LC3B
(clone 2G6, diluted 1:500 for immunoblotting or 1:100 for immunofluorescence) is
available from NanoTools, Teningen, Germany. The mouse monoclonal anti-p62 antibody
(diluted 1:1000 to 1:5000 for immunoblotting or 1:100 for immunofluorescence) was
purchased from BD Transduction Laboratories, Franklin Lakes, NJ, USA (order number
610832). The polyclonal BacTrace goat anti-Salmonella CSA-1 antibody
(diluted 1:200) was purchased from KPL, Gaithersburg, MD, USA (order number
01-91-99). The mouse anti-GAPDH (clone GAPDH-71.1, diluted 1:50,000) is available
from Sigma. The rabbit anti-ubiquitin serum is available from Sigma. Secondary
antibodies for immunofluorescence were Alexa Fluor 488 or 546-conjugated goat
anti-mouse IgG (diluted 1:1000) from Invitrogen, Alexa Fluor 647-conjugated goat
anti-rabbit IgG (diluted 1:500) and CyTM5-conjugated AffiniPure donkey
anti-goat IgG (diluted 1:400) from Jackson ImmunoResearch Laboratories, West Grove,
PA, USA and Alexa Fluor 405-conjugated donkey anti goat IgG (diluted 1:200) from
Abcam, Cambridge, UK.
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2

Western Blot Analysis of Key Proteins

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Cells were lysed with sample loading buffer (2% w/v sodium dodecyl sulfate (SDS), 62.5 mM Tris-HCl, 10% v/v glycerol, 2.5% v/v 2-mercaptoethanol, 0.005% w/v bromophenol blue) to obtain total cell lysate samples, which were separated by SDS-PAGE using ClearPAGE 4–20% polyacrylamide gradient gels (Expedeon) and transferred onto an Immobilon-FL PVDF membrane (EMD Millipore). Membranes were then blocked with Odyssey Blocking Buffer (Li-Cor) before incubation with primary antibodies for 1 hour at room temperature and subsequent incubation with IRDyes-conjugated secondary antibodies (Li-Cor) for 1 hour at room temperature. Mouse anti-GAPDH (clone GAPDH-71.1) and rabbit anti-GAPDH (G9545) were from Sigma-Aldrich. IRDye 800CW-conjugated streptavidin was from Li-Cor, while the anti-Lck (clone L22B1), anti-p44/42 MAPK (Erk1/2, clone 3A7), and anti-phospho-p44/42 MAPK (Erk1/2, Thr202/Tyr204, clone D13.14.4E) antibodies were from Cell Signaling Technologies. Blots were visualized using the Odyssey CLx Imaging System and quantified using ImageStudio software (Li-Cor).
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