Application
Western blot
Sample
Mouse Tissue lysate - whole (Ms Brain Ms Heart Ms Spleen Raw264.7)
Loading amount
20 µg
Specification
Ms Brain Ms Heart Ms Spleen Raw264.7
Gel Running Conditions
Reduced Denaturing (Pierce precise protein gel 8-16%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Other product details
Concentration
1 µg/ml
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 2.5% Non-fat Milk, PBS-Tween 0.1%
Secondary antibody
Name
Non-Abcam antibody was used: Swine anti-Rabbit HRP
Host species: Pig
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Pig
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
Supersignal West dura
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 60 kDa Non-specific: 43 kDa
Additional data
Additional Notes
Lane order (1-4): Ms Brain Ms Heart Ms Spleen Raw264.7
A species of ~60 kDa close to the predicted mobility of 52 kDa was detected in all of the murine lysates tested. Note other commercial antibodies identify Meis2 as a 60 kDa species (scbt meis2-n-17-antibody) An additional, faster migrating (~43 kDa) species was observed in Ms Heart lysate, but the origin of this species is not clear. Note, this antibody will not recognise the shorter isoforms of Meis2 as the epitope is raised to the C-terminus, which is spliced out in isoforms A and B (Swissprot: P97367).
A species of ~60 kDa close to the predicted mobility of 52 kDa was detected in all of the murine lysates tested. Note other commercial antibodies identify Meis2 as a 60 kDa species (scbt meis2-n-17-antibody) An additional, faster migrating (~43 kDa) species was observed in Ms Heart lysate, but the origin of this species is not clear. Note, this antibody will not recognise the shorter isoforms of Meis2 as the epitope is raised to the C-terminus, which is spliced out in isoforms A and B (Swissprot: P97367).
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.
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投稿 May 25 2010