Application
Western blot
Sample
Bacteria Cell lysate - whole cell (Sinorhizobium meliloti whole cell lysate)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
3e+007 cells
Treatment
in SDS sample buffer 2-mercaptoethanol
Specification
Sinorhizobium meliloti whole cell lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: TBST (Tris buffered saline and Tween-20)
Concentration
0.005 µg/ml
Secondary antibody
Dilution
1/25000
Name
Non-Abcam antibody was used: Donkey anti-mouse IgG
Host species: Donkey
Clonality: Monoclonal
Conjugation: HRP polymer
Host species: Donkey
Clonality: Monoclonal
Conjugation: HRP polymer
Detection
Detection method
ECL+
Exposure
30 second(s)
Bands
Specific: 55 kDa
Positive control
Wild-type Sinorhizobium meliloti strain Rm1021, 1/2 dilution, and 1/5 dilution
Additional data
Additional Notes
wet transfer onto PVDF membrane overnight at 15 V, chemiluminescence reagent used capable of detecting femtograms of proteins, surprisingly low concentrations of primary antibody (20 mL of 5 ng antibody / mL TBST, for a total of 100 ng antibody per blot) was successfully for detecting single band, recommended dilution of primary antibody (1 ug/mL) led to extremely high signal and background
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.
Joseph Chen
Verified customer
投稿 Mar 06 2017