Application
Western blot
Sample
Frog Tissue lysate - whole (frog brain: forebrain, midbrain, hindbrain)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
frog brain: forebrain, midbrain, hindbrain
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 23°C
Other product details
Incubation time
20 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS-T
Dilution
1/1000
Secondary antibody
Dilution
1/3000
Name
Non-Abcam antibody was used: BioRad Goat Anti-Mouse IgG (H+L) Horseradish Perox
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
5 second(s)
Positive control
15 ug of mouse brain homogenate (hippocampus - lane 11 brainstem - lane 12) was used as a loading control (observed bands at ~100 kDa as expected)
Additional data
Additional Notes
ab2826 was predicted to work in amphibian (species of interest), however this antibody did not work with my frog brain homogenates. Worked as expected with mouse brain samples.
For WB, BioRad Mini-PROTEAN® TGX Stain-Free™ Precast Gels (7.5%) were used.
Lanes on image:
1- marker (Fisher BioReagents™ EZ-Run™ Prestained Rec Protein Ladder, Fisher BioReagents). Molecular weights: ~170 kDa, ~130, ~95, ~72, ~55, ~43, ~34, ~26, ~17, ~10.
2- 30 ug midbrain
3- 40 ug midbrain
4- 50 ug midbrain
5- 30 ug hindbrain
6- 40 ug hindbrain
7- 50 ug hindbrain
8- 30 ug forebrain
9- 40 ug forebrain
10- 50 ug forebrain
11- 15 ug hippocampus (mouse)
12- 15 ug brainstem (mouse)
Abcam response
Abcam has not tested ab2826 in Frog samples and thus could not guarantee the antibody performance in this species. This individual received a credit for testing the antibody in Frog samples as part of our AbTrial program.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Abcam user community
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提交于 Mar 07 2019