Anti-Chk1 antibody [EP691Y] - BSA and Azide free

6 product images

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  Immunocytochemistry/ Immunofluorescence - Anti-Chk1 antibody [EP691Y] - BSA and Azide free (ab210964)

  Clone EP691Y (ab210964) has been successfully conjugated by Abcam. This image was generated using Anti-Chk1 antibody [EP691Y] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-Chk1 antibody [EP691Y] ab196520 for protocol details.

  Alexa Fluor® 647 Anti-Chk1 antibody [EP691Y] ab196520 staining Chk1 in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Alexa Fluor® 647 Anti-Chk1 antibody [EP691Y] ab196520 at 1/100 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 488, shown in green) at 1/167 dilution, overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Immunocytochemistry/ Immunofluorescence - Anti-Chk1 antibody [EP691Y] - BSA and Azide free (ab210964)

  Clone EP691Y (ab210964) has been successfully conjugated by Abcam. This image was generated using Anti-Chk1 antibody [EP691Y] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-Chk1 antibody [EP691Y] ab196308 for protocol details.

  Alexa Fluor® 488 Anti-Chk1 antibody [EP691Y] ab196308 staining Chk1 in MCF-7cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with Alexa Fluor® 488 Anti-Chk1 antibody [EP691Y] ab196308 at 1/500 dilution (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 594, shown in red) at 1/167 dilution overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Flow Cytometry (Intracellular) - Anti-Chk1 antibody [EP691Y] - BSA and Azide free (ab210964)

  Overlay histogram showing HeLa cells stained with Anti-Chk1 antibody [EP691Y] ab40866 (red line). The cells were fixed with 80% methanol (5 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 followed by the antibody (Anti-Chk1 antibody [EP691Y] ab40866, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Chk1 antibody [EP691Y] ab40866).
  

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  Immunocytochemistry/ Immunofluorescence - Anti-Chk1 antibody [EP691Y] - BSA and Azide free (ab210964)

  Immunofluorescence staining of MCF7 cells with purified Anti-Chk1 antibody [EP691Y] ab40866 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was an Alexa Fluor^® 488 conjugated goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Chk1 antibody [EP691Y] ab40866).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chk1 antibody [EP691Y] - BSA and Azide free (ab210964)

  Immunohistochemical analysis of Chk1 expression in paraffin-embedded human breast carcinoma using Anti-Chk1 antibody [EP691Y] ab40866 at a 1:250 dilution. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Chk1 antibody [EP691Y] ab40866).

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  Western blot - Anti-Chk1 antibody [EP691Y] - BSA and Azide free (ab210964)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Chk1 antibody [EP691Y] ab40866).
  
  Anti-CHEK1 antibody [EP691Y] (Anti-Chk1 antibody [EP691Y] ab40866) staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-Chk1 antibody [EP691Y] ab40866 was shown to bind specifically to CHEK1. A band was observed at 57 kDa in wild-type A549 cell lysates with a reduction in signal observed at this size in CHEK1 heterozygous knockout cell line. To generate this image, wild-type and CHEK1 heterozygous knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Chk1 antibody [EP691Y] (Anti-Chk1 antibody [EP691Y] ab40866) at 1/10000 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: Western blot - Human CHEK1 heterozygous knockout A549 cell line (Human CHEK1 heterozygous knockout A549 cell line ab276102)

  Lane 2: CHEK1 knockout A549 cell lysate at 20 µg

  Lane 3: A431 cell lysate at 20 µg

  Lane 4: MOLT-4 cell lysate at 20 µg

  Secondary

  Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 54 kDa

  Observed band size: 57 kDa