Anti-6X His tag® antibody [4D11] (ab5000)

Western blot - 6X His tag® antibody [4D11] (ab5000)

All lanes : Anti-6X His tag® antibody [4D11] (ab5000) at 1/2000 dilution

Lane 1 : Lysates prepared from HEK-293 cells
Lane 2 : Lysates prepared from HEK-293 cells transfected with Beta3-His (CACNB3)

Secondary
HRP-conjugated goat anit-mouse IgG polyclonal at 1/5000 dilution

This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic

Immunoprecipitation - 6X His tag® antibody [4D11] (ab5000)

ab5000 Immunoprecipitate in human HEK293 whole cell lysate. 25µg of cell lysate incubated with primary antibody (undiluted) and matrix (Protein G) for 16 hours at 4°C. For western blotting an undiluted HRP-Goat anti-rabbit IgG polyclonal was used. Beta3-His fusion protein was immunoprecipitated using anti His Ab, and specific beta3a band (56kDa) was detected using anti beta3 Ab

Lane 1. Lysate of HEK293 cells expressing beta3-His (CACNB3)

Lane 2. IP with anti His Ab

Lane 3. Non bound fraction

This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic

Western blot - Anti-6X His tag® antibody [4D11] (ab5000)

Anti-6X His tag® antibody [4D11] (ab5000) at 1/1000 dilution + 6X His Tagged Protein at 1 µg

Secondary
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/3000 dilution
developed using the ECL technique

Performed under reducing conditions.

Exposure time : 30 seconds

Immunocytochemistry/ Immunofluorescence - 6X His tag® antibody [4D11] (ab5000)

ab5000 staining Human HEK 293 cells transfected with CACNB3-His by ICC/IF. Cells were PFA fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 5% serum for 20 minutes at 25°C. The primary antibody (1/250 in 1% goat serum, 0.1% TX100, 1X PBS) was incubated with the sample for 16 hours at 4°C. An Alexa Fluor^® 546 conjugated goat anti-mouse antibody (1/500) was used as the secondary.  DAPI was used for nuclei staining (blue).

This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic