Mouse IgG2c, kappa monoclonal [18C8BC7AD10] - Isotype Control

• Flow Cyt

Supplier Data

Flow Cytometry - Mouse IgG2c, kappa monoclonal [18C8BC7AD10] - Isotype Control (AB170191)

Flow cytometry staining of C57 BL/6 mouse bone marrow cells with ab320708 (right) or Mouse IgG2c, kappa monoclonal (ab170191), isotype control (left). Cells were incubated for 30 min on ice in 1x PBS containing 10%rat serum 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab320708 or Mouse IgG2c, kappa monoclonal (ab170191), isotype control (1x 10e6 in 100 μl at 1.0 μg/ml) for 30min on ice. The cells were simultaneously stained with APC anti-mouse Ly-6C Antibody.

The secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.

• IP

Lab

Immunoprecipitation - Mouse IgG2c, kappa monoclonal [18C8BC7AD10] - Isotype Control (AB170191)

TATA binding protein TBP was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate (10 µg) with ab300656 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300656 at 1/2000 dilution. Mouse IgG for IP (HRP) (ab131368) was used as the secondary at 1/5000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-TATA binding protein TBP antibody [mAbcam51841] - Loading Control (<a href='/en-us/products/primary-antibodies/tata-binding-protein-tbp-antibody-mabcam51841-loading-control-ab300656'>ab300656</a>) at 1/30 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

HeLa whole cell lysate

Secondary

All lanes:

Immunoprecipitation - Anti-mouse IgG for IP (HRP) (<a href='/en-us/products/secondary-antibodies/mouse-igg-for-ip-hrp-ab131368'>ab131368</a>) at 1/5000 dilution

Observed band size: 35 kDa,38 kDa

false

Exposure time: 24s

• IP

Lab

Immunoprecipitation - Mouse IgG2c, kappa monoclonal [18C8BC7AD10] - Isotype Control (AB170191)

TATA binding protein TBP was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate (10 µg) with ab300656 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300656 at 1/2000 dilution. Mouse IgG for IP (HRP) (ab131368) was used as the secondary at 1/5000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-TATA binding protein TBP antibody [mAbcam51841] - Loading Control (<a href='/en-us/products/primary-antibodies/tata-binding-protein-tbp-antibody-mabcam51841-loading-control-ab300656'>ab300656</a>) at 1/30 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 10 µg

Lane 2:

NIH/3T3 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - Anti-mouse IgG for IP (HRP) (<a href='/en-us/products/secondary-antibodies/mouse-igg-for-ip-hrp-ab131368'>ab131368</a>) at 1/5000 dilution

Predicted band size: 38 kDa

Observed band size: 36 kDa

false

Exposure time: 24s