Western blot abreview for Anti-alpha 3 Sodium Potassium ATPase antibody [XVIF9-G10]

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Abreviews
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

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.

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Submitted Mar 07 2019

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