Application
Western blot
Sample
Human Cell lysate - whole cell (Human Embryonic Stem cells)
Loading amount
70000 cells
Specification
Human Embryonic Stem cells
Gel Running Conditions
Reduced Denaturing (10% SDS)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Other product details
Dilution
1/400
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% BSA in TBS-T
Secondary antibody
Name
Non-Abcam antibody was used: DAKO: Polyclonal Goat anti Rabbit HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer
Dilution
1/2000
Detection
Detection method
ECL Advance
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 180, 90 kDa Non-specific: 60 kDa
Positive control
Lane 1: HUES7 cells
Lane 2: H9 cells
Lane 3: iPS cells
Lane 2: H9 cells
Lane 3: iPS cells
Additional data
Additional Notes
1% BSA blocking solution resulted in much clearer bands than either 5% BSA or 5% milk blocking solution.
Exposure time more than 1 minute gives very high background noise.
Exposure time more than 1 minute gives very high background noise.
Abcam response
Thank you for sharing this data with us. ab29112 has previously been tested on mouse ES cells, where we see ~140kDa and ~85kDa bands, which we believe correspond to the two Sall4 isoforms. Ma et al, 2006 (PMID: 16763212) have shown these to run even higher than predicted for human Sall4a (165kDa) and Sall4b (95kDa). We believe that these are the top 2 bands seen here. Abcam would welcome feedback from other customers using this product.
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
Verified customer
提交于 Dec 12 2012