Application
Western blot
Sample
Mouse Cell lysate - whole cell (Lysates of BMDCs)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
25000 cells
Treatment
LPS (3h, 50 ng/ml) +/- Nigericin (5 µM, 2.5h)
Specification
Lysates of BMDCs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 2% SMP in PBS/T
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L)
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer
Detection
Detection method
ECL
Negative control
Lane 3 and 4, BMDCs of GSDMD -/-
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 53, 32 kDa
Positive control
Lane 1 and 2, BMDCs of WT, +/- Nigericin
Additional data
Additional Notes
Very good antibody that shows strong and GSDMD-specific bands. Detects both, full lenght as well as cleaved GSDMD.
Detects cleaved GSDMD better than the full lenght protein. (Lane 1) WT, LPS (Lane 2) WT, LPS+Nigericin, (Lane 3) GSDMD ko, LPS and (Lane 4) GSDMD ko, LPS, Nigericin.
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Abcam user community
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Submitted Aug 23 2020