Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free

38 product images

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  Image from Simeone P et al. PLoS One. 2014;;9(7):e103030.; doi: 10.1371/journal.pone.0103030.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Expression of c-Myc, as determined by immunohistochemical staining of glioblastoma sample (left) and low-grade glioma tumor (right) with Anti-c-Myc antibody [Y69] - ChIP Grade ab32072. Representative samples are shown. Scale bars = 20 μm. Nuclei were counterstained with hematoxylin (in blue).
  

  For the full image see PMID 25050814.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Image from Toon CW et al. PLoS One. 2014;9(2):e87456. Fig 1.; doi: 10.1371/journal.pone.0087456.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunohistochemistry staining of colorectal carcinoma using rabbit anti-c-Myc antibody
  
  Human colorectal carcinoma (CRC) tissues stained for c-Myc using Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at 1/100 dilution in immunohistochemical analysis.

  Panel A: c-Myc positive IHC staining; Panel B: c-Myc negative IHC staining.

  For the full image see PMID 24503701.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Image from Kluk MJ et al. PLoS One. 2012;7(4):e33813. Fig 1.; doi: 10.1371/journal.pone.0033813.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Photomicrographs of select tumors and reactive tissue stained for c-Myc (positive staining = brown nuclei). Positive control (Burkitt lymphoma with a confirmed c-Myc translocation) revealed uniform, intense staining in >90% of tumor cells (Burkitt). In contrast, reactive lymphoid tissue revealed variable staining in only 10% of normal lymphocyte nuclei (Tonsil). Representative images from Diffuse large B cell lymphoma (DLBCL) cases and associated percent c-Myc+ tumor nuclei: Case 1, 90% MYC+; Case 7, 70% MYC+; and Cases 35 and 38, 30% c-Myc+. c-Myc staining was exclusively nuclear in all cases under the described staining conditions.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunoprecipitation - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc was immunoprecipitated using 0.5mg Jurkat (human T cell leukemia cell line from peripheral blood) whole cell extract, 5µg of unpurified rabbit monoclonal to c-Myc [Y69] 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, Jurkat 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°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with unpurified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072.

  Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

  Band: 57kDa; c-Myc [Y69]

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

  All lanes: Immunoprecipitation - Anti-c-Myc antibody [Y69] - ChIP Grade (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072)

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 48 kDa

  Exposure time: 12min

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  Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  False colour image of Western blot: Anti-c-Myc antibody [Y69] staining at 1/1000 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-c-Myc antibody [Y69] - ChIP Grade ab32072 was shown to bind specifically to c-Myc. A band was observed at 45/57 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in MYC CRISPR-Cas9 edited cell line Human MYC (c-Myc) knockout HEK-293T cell line ab256500 (CRISPR-Cas9 edited cell lysate Human MYC (c-Myc) knockout HEK-293T cell lysate ab263850). The band observed in the CRISPR-Cas9 edited lysate lane below 45/57 kDa is likely to represent a truncated form of c-Myc. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MYC CRISPR-Cas9 edited HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

  All lanes: Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072) at 1/1000 dilution

  Lane 1: Wild-type HEK-293T cell lysate at 20 µg

  Lane 2: MYC CRISPR-Cas9 edited HEK-293T cell lysate at 20 µg

  Lane 2: Western blot - Human MYC (c-Myc) knockout HEK-293T cell line (Human MYC (c-Myc) knockout HEK-293T cell line ab256500)

  Lane 3: Jurkat cell lysate at 20 µg

  Lane 4: SH-SY5Y cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 48 kDa

  Observed band size: 45 kDa, 57 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072). Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 staining MYC in wild-type HEK293 cells (top panel) and MYC knockout HEK293 cells (Human MYC (c-Myc) knockout HEK-293T cell line ab256500) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at 5μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
  Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

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  Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Blocking buffer and concentration: 5% NFDM/TBST.
  

  Diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  All lanes: Raji (human Burkitt's lymphoma) whole cell lysate at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051)

  Predicted band size: 48 kDa

  Observed band size: 57 kDa

  Exposure time: 10s

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  Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  This data was developed using Anti-c-Myc antibody [Y69] - ChIP Grade ab32072, the same antibody clone in a different buffer formulation. Different batches of Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 were tested on Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 0.2 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 57 kDa.

  All lanes: Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072)

  Predicted band size: 48 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Clone Y69 (ab168727) has been successfully conjugated by Abcam. This image was generated using Anti-c-Myc antibody [Y69] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-c-Myc antibody [Y69] ab190560 for protocol details.

  Alexa Fluor® 647 Anti-c-Myc antibody [Y69] ab190560 staining c-Myc in panc1 cells. The cells were fixed with 100% methanol (5 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-c-Myc antibody [Y69] ab190560 at a working dilution of 1 in 100 (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 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

  This product also gave a positive signal in 4% formaldehyde (10 min) fixed panc1 cells under the same testing conditions.

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

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  Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Clone Y69 (ab168727) has been successfully conjugated by Abcam. This image was generated using Anti-c-Myc antibody [Y69] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-c-Myc antibody [Y69] ab190026 for protocol details.

  Alexa Fluor® 488 Anti-c-Myc antibody [Y69] ab190026 staining c-myc in Panc-1 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Alexa Fluor® 488 Anti-c-Myc antibody [Y69] ab190026 at 1/100 dilution (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

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

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  Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunofluorescence staining of HeLa cells using rabbit anti-c-Myc antibody
  
  Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 staining c-Myc in HeLa (human epithelial cell line from cervix adenocarcinoma) cells. The cells were fixed with 4% formaldehyde (10min) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at 10μg/ml dilution (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594) at 2μg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

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

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunofluorescence staining of HeLa cells using rabbit anti-c-Myc antibody
  
  Immunocytochemistry/immunofluorescence analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling c-Myc with purified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at 1/100 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody (1/500 dilution) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

  Control: primary antibody (1/100 dilution) and secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) (1/500 dilution).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Flow Cytometry (Intracellular) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc flow cytometry staining of HeLa cells using rabbit anti-c-Myc antibody
  
  Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 (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-c-Myc antibody [Y69] - ChIP Grade ab32072 1/76 dilution) for 30 min at 22°C. The secondary antibody used wasGoat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibodyat 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG [EPR25A] (monoclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

  Acquisition of >5000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 nm bandpass filter.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling c-Myc with Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution. Nuclear staining on mouse spleen. Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunohistochemistry staining of human esophagus using rabbit anti-c-Myc antibody
  
  IHC image of Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 staining c-Myc in human esophagus formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. 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 Anti-c-Myc antibody [Y69] - ChIP Grade ab32072, 1µg/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. No primary antibody was used in the Secondary only control (shown on the inset).

  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.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Image courtesy of Dr Vladimir Milenkovic by customer review.

  Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunofluorescence staining of HEK293 cells using rabbit anti-c-Myc antibody
  
  Unpurified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 staining c-Myc in HEK293 cells transfected with CACNB4-c-Myc by immunocytochemistry/ immunofluorescence.Cells were fixed in paraformaldehyde permeabilized with 0.5% Triton X-100 then blocked using 5% serum for 20 minutes at 25°C. Samples were then incubated with Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at a 1/250 dilution for 16 hours at 4°C. The secondary used was an Alexa Fluor^® 488 conjugated goat anti-rabbit polyclonal used at a 1/500 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunohistochemistry staining of human adenocarcinoa using rabbit anti-c-Myc antibody
  
  IHC image of Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 staining c-Myc in human adenocarcinoma formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 Anti-c-Myc antibody [Y69] - ChIP Grade ab32072, 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. No primary antibody was used in the negative control (shown on the inset).

  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.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunohistochemistry staining of adenocarcinoma using rabbit anti-c-Myc antibody
  
  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of the colon tissue labelling c-Myc with purified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at 1/500 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody was used as the secondary antibody (1/500 dilution). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human diffuse large B cell lymphoma tissue labelling c-Myc with purified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skin carcinoma tissue labelling c-Myc with unpurified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at 1/50.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunofluorescence staining of HeLa cells using rabbit anti-c-Myc antibody
  
  ICC/IF image of unpurified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 stained HeLa (human epithelial cell line from cervix adenocarcinoma) cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 1μg/ml) overnight at +4°C. The secondary antibody (green) was Goat Anti-Rabbit IgG H&L (DyLight^® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) used at a 1/250 dilution for 1h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunohistochemistry staining of colon tissue using rabbit anti-c-Myc antibody
  
  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarinoma of colon tissue labelling c-Myc with unpurified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human urinary bladder transitional carcinoma tissue labelling c-Myc with unpurified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072.
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunohistochemistry staining of lung adenocarcinoma using rabbit anti-c-Myc antibody
  
  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung adenocarinoma tissue labelling c-Myc with unpurified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric adenocarcinoma tissue labelling c-Myc with unpurified Anti-c-Myc antibody [Y69] - ChIP Grade ab32072.
  
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  OI-RD Scanning - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
  Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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  Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Blocking and dilution buffer: 5% NFDM/TBST.
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

  All lanes: Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072) at 1/1000 dilution

  Lane 1: MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 2: Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 20 µg

  Lane 3: K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates at 20 µg

  Lane 4: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates at 20 µg

  Lane 5: THP-1 (Human monocytic leukemia monocyte) whole cell lysates at 20 µg

  Lane 6: Rat spleen whole cell lysates at 20 µg

  Lane 7: L6 (Rat skeletal muscle myoblast) whole cell lysates at 20 µg

  Lane 8: Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates at 20 µg

  Lane 9: RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 20 µg

  Secondary

  All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/20000 dilution

  Predicted band size: 48 kDa

  Observed band size: 57 kDa, 48 kDa, 45 kDa

  Exposure time: 3min

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunohistochemistry staining of human colon using rabbit anti-c-Myc antibody
  
  

  This data was developed using the same antibody clone in a different buffer formulation (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

  Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling c-Myc with Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at a concentration of 1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 anti-c-Myc antibody [Y69] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  c-Myc immunohistochemistry staining of human colon using rabbit anti-c-Myc antibody
  
  

  This data was developed using the same antibody clone in a different buffer formulation (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

  Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling c-Myc with Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at a concentration of 1 µg/ml. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins. Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 anti-c-Myc antibody [Y69] was incubated for 30 mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.

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  Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

  Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 staining of Myc proto-oncogene protein in a HCT 116 cell spheroid. The cells were fixed with 4% paraformaldehyde (10 min), permeabilised with 2% Triton X-100 for 1h and then blocked with 1% BSA/10% normal goat serum/0.3M glycine 0.1% PBS-Tween overnight at room temperature. The spheroids were then incubated overnight at room temperature with Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at 2 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 2 μg/ml. DAPI was used as nuclear counterstain (shown in blue). As secondary antibodies Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat anti-Rabbit (Alexa Fluor^® 488) (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat anti-Mouse (Alexa Fluor^® 594) (shown in magenta) were used, incubated overnight at room temperature. All permeabilization, blocking and antibody incubation steps were performed using a rotary shaker.

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

  Similar results were observed using 100% Methanol (5min).

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  ChIC/CUT&RUN sequencing - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 5 µg of Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 [Y69]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.
  
  Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section.
  
  The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
  
  This data was developed using the same antibody clone in a different buffer formulation (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

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  Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
  Lanes 1 and 2: 80 seconds exposure time.
  Lanes 3 to 5: 5 seconds exposure time.
  
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

  All lanes: Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072) at 1/1000 dilution

  Lane 1: Rat pancreas lysates at 20 µg

  Lane 2: AR42J (Rat pancreatic tumor epithelial cell) whole cell lysates at 20 µg

  Lane 3: Rat-1 (Rat embryonic fibroblast) whole cell lysates at 20 µg

  Lane 4: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

  Lane 5: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 20000 µg/mL

  Observed band size: 45,57 kDa

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  ChIP-sequencing - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HeLa cells and 8µg of Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 [Y69]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
  Additional screenshots of mapped reads can be downloaded here.
  This data was developed using the same antibody clone in a different buffer formulation (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

• 

  Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Blocking buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072) at 1/1000 dilution

  Lane 1: Mouse brain tissue lysate at 20 µg

  Lane 2: Mouse brain cancer tissue lysate at 20 µg

  Lane 3: Mouse skin tissue lysate at 20 µg

  Lane 4: Mouse skin cancer tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 45 kDa

  Exposure time: 60s

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  Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Blocking buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-c-Myc antibody [Y69] - ChIP Grade (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072) at 1/1000 dilution

  Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 4: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 5: A549 (Human lung carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 6: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 45 kDa, 57 kDa

  Exposure time: 20s

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  ChIP-sequencing - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 NCCIT cells and 8µg of Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 [Y69]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
  Additional screenshots of mapped reads can be downloaded here.
  This data was developed using the same antibody clone in a different buffer formulation (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

• 

  ChIP-sequencing - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 NCCIT cells and 8µg of Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 [Y69]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
  Additional screenshots of mapped reads can be downloaded here.
  This data was developed using the same antibody clone in a different buffer formulation (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).

• 

  ChIP-sequencing - Anti-c-Myc antibody [Y69] - ChIP Grade - BSA and Azide free (ab168727)

  Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 NCCIT cells and 8µg of Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 [Y69]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
  Additional screenshots of mapped reads can be downloaded here.
  This data was developed using the same antibody clone in a different buffer formulation (Anti-c-Myc antibody [Y69] - ChIP Grade ab32072).