Western blot abreview for Anti-GnRH antibody

Good
Abreviews
Application
Western blot
Sample
Mouse Recombinant protein (mixed with LbT2 cells 10ug lysate)
Gel Running Conditions
Reduced Denaturing (12.5%)
Loading amount
0.054 µg
Specification
mixed with LbT2 cells 10ug lysate
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Other product details

Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBST
Dilution
1/1000

Secondary antibody

Dilution
1/10000
Name
Non-Abcam antibody was used: goat anti rabbit HRP conjugated
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer

Detection

Detection method
ECL
Negative control
The LbT2 cell lysate not mixed with the recombinant protein
Exposure
30 second(s)
Bands
Specific: 47 kDa Non-specific: 15 kDa
Positive control
The recombinant protein alone

Additional data

Additional Notes
GnRH lys6 was cross linked to a GFP tagged protein (471 amino acids). The protein is a chimeric protein, not yet published. The sequence of the GnRH lys6 is : EHWSYKLRPG. the cross linker is sulfo-GMBS, which cross links the lys6 of the GnRH to a cysteine in the chimeric protein. The cross linking was verified by mass spec. Lanes: 1) LbT2 cell lysate 10ug . 2) LbT2 cell lysate 10ug + 54ng of the GnRH lys6 cross linked protein. 3) LbT2 cell lysate 10ug + 5.4ng of the GnRH lys6 cross linked protein. 4) LbT2 cell lysate 10ug + 0.54ng of the GnRH lys6 cross linked protein. 5) 54ng of the GnRH lys6 cross linked protein 6) pre-stained marker lane.

Dr. Lilach Polinsky

Verified customer

Submitted Oct 10 2018

Sign up