Anti-RAD54抗体

• IP

Unknown

Immunoprecipitation - Anti-RAD54 antibody (AB10705)

RAD54 was immunoprecipitated using 0.5mg Hela whole cell extract, 5μg of Rabbit polyclonal to RAD54 and 50μl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70^oC; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab10705.
Secondary : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band : 70kDa; RAD54, non specific band : 75 kDa, We are unsure as to the identity of this extra band. 99kDa band : due to background seen in No ab control lane (2).

All lanes:

Immunoprecipitation - Anti-RAD54 antibody (ab10705)

Predicted band size: 84 kDa

false

• WB

Unknown

Western blot - Anti-RAD54 antibody (AB10705)

Rabbit polyclonal to RAD54 (ab10705) at 1/500.

Lane 1 Marker
Lane 2 HeLa nuclear extract
Lane 3 HeLa cell lysate
Lane 4 A432 cell lysate
Lane 5 MCF7 cell lysate
Lane 6 HEK 293 cell lysate

Secondary antibody - Goat polyclonal to rabbit IgG (HRP) - ab6721.

Lane 1:

Western blot - Anti-RAD54 antibody (ab10705)

Lanes 2 - 6:

Western blot - Anti-RAD54 antibody (ab10705) at 1/500 dilution

Lane 1:

Marker

Lane 2:

HeLa nuclear cell lysate

Lane 3:

HeLa cell lysate

Lane 4:

A432 cell lysate

Lane 5:

MCF7 cell lysate

Lane 6:

Hek 293 cell lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab6721'>ab6721</a>)

Predicted band size: 84 kDa

Observed band size: 70 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAD54 antibody (AB10705)

IHC image of ab10705 staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab10705, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.