Rat IgG2a, kappa monoclonal [RTK2758] - Isotype Control

• Flow Cyt

Unknown

Flow Cytometry - Rat IgG2a, kappa monoclonal [RTK2758] - Isotype Control (AB18450)

Overlay histogram showing HeLa cells stained with ab53358 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53358, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rat IgG2a, kappa monoclonal [aRTK2758] (ab18450, 2μg/1x10⁶ cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

• IP

Lab

Immunoprecipitation - Rat IgG2a, kappa monoclonal [RTK2758] - Isotype Control (AB18450)

Serum Response Factor SRF was immunoprecipitated from HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate with ab252868 at 1/30 dilution (2μg in 0.35mg lysates) (36.067μg/ml). Western blot was performed on the immunoprecipitate using ab252868 at 1/1000 dilution. Goat Anti-rat IgG (H+L), HRP) (ab205720) was used at 1/10000 dilution.

Lane 1 : HeLa whole cell lysate 10μg.

Lane 2 : ab252868 IP in HeLa whole cell lysate.

Lane 3 : Rat monoclonal IgG2a (ab18450) instead of ab252868 in HeLa whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 10 seconds.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11893590and PMID : 10642500).

All lanes:

Immunoprecipitation - Anti-Serum Response Factor SRF antibody [2C5] (<a href='/en-us/products/primary-antibodies/serum-response-factor-srf-antibody-2c5-ab252868'>ab252868</a>) at 1/1000 dilution

Predicted band size: 52 kDa

Observed band size: 57 kDa,67 kDa

false

Exposure time: 6s

• Flow Cyt

Supplier Data

Flow Cytometry - Rat IgG2a, kappa monoclonal [RTK2758] - Isotype Control (AB18450)

Flow cytometric analysis of A20 (mouse reticulum sarcoma B lymphocyte) cells labelling Transferrin Receptor with ab256568 at 1/500 dilution (0.1μg) (Red) compared with a Rat monoclonal IgG2a (ab18450) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat F(ab)2 Anti-Rat IgG Fc (Alexa Fluor^® 488, ab150161) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Rat IgG2a, kappa monoclonal [RTK2758] - Isotype Control (AB18450)

Flow cytometry staining of C57 BL/6 mouse bone marrow cells with ab320706 (right) or Rat IgG2a, kappa monoclonal (ab18450), isotype control (left). Cells were incubated for 30 min on ice in 1x PBS containing 10% mouse serum and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab320706 or Rat IgG2a, kappa monoclonal (ab18450), isotype control (1x 106 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-Rat 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 - Rat IgG2a, kappa monoclonal [RTK2758] - Isotype Control (AB18450)

Serum Response Factor SRF was immunoprecipitated from C2C12 (mouse myoblasts myoblast), whole cell lysate with ab252868 at 1/30 dilution (2μg in 0.35mg lysates) (36.067μg/ml). Western blot was performed on the immunoprecipitate using ab252868 at 1/1000 dilution. Goat Anti-rat IgG (H+L), HRP) (ab205720) was used at 1/10000 dilution.

Lane 1 : C2C12 whole cell lysate 10μg.

Lane 2 : ab252868 IP in C2C12 whole cell lysate.

Lane 3 : Rat monoclonal IgG2a (ab18450) instead of ab252868 in C2C12 whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 6 seconds.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11893590 and PMID : 10642500).

All lanes:

Immunoprecipitation - Anti-Serum Response Factor SRF antibody [2C5] (<a href='/en-us/products/primary-antibodies/serum-response-factor-srf-antibody-2c5-ab252868'>ab252868</a>) at 1/1000 dilution

Predicted band size: 52 kDa

Observed band size: 57 kDa,67 kDa

false

Exposure time: 6s