Anti-c-Myc antibody [Y69]
| Name | Anti-c-Myc antibody [Y69] |
|---|---|
| Supplier | Abcam |
| Catalog | ab32072 |
| Prices | $403.00 |
| Sizes | 100 µl |
| Host | Rabbit |
| Clonality | Monoclonal |
| Isotype | IgG |
| Clone | Y69 |
| Applications | WB IHC-P ICC/IF ICC/IF IP |
| Species Reactivities | Mouse, Rat, Human |
| Antigen | Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human c-Myc aa 1-100 (N terminal) |
| Blocking Peptide | c-Myc peptide |
| Description | Rabbit Monoclonal |
| Gene | MYC |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
ab32072 staining c-Myc in HeLa 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 ab32072 at 10μg/ml dilution (shown in green) and 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).
IHC image of 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 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.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling c-Myc with purified ab32072 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.Control: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Unpurified 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 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.See Abreview
ICC/IF image of unpurified ab32072 stained HeLa 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 (ab32072, 1µg/ml) overnight at +4°C. The secondary antibody (green) was anti rabbit DyLight® 488 IgG - H&L, pre-adsorbed (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.
![All lanes : Anti-c-Myc antibody [Y69] (ab32072) at 1/10000 dilution (purified)Lane 1 : Raji cell lysateLane 2 : K562 cell lysateLane 3 : Jurkat cell lysateLysates/proteins at 20 µg per lane.SecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/16497_ab32072-239306-ab32072wb.jpg)
All lanes : Anti-c-Myc antibody [Y69] (ab32072) at 1/10000 dilution (purified)Lane 1 : Raji cell lysateLane 2 : K562 cell lysateLane 3 : Jurkat cell lysateLysates/proteins at 20 µg per lane.SecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
![All lanes : Anti-c-Myc antibody [Y69] (ab32072) at 1/1000 dilution (unpurified)Lane 1 : Raji (Human Burkitt's lymphoma cell line) Whole Cell LysateLane 2 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell LysateLane 3 : THP1 (Human acute monocytic leukemia cell line) Whole Cell LysateLane 4 : A20 (Mouse B lymphoma cell line) Whole Cell LysateLane 5 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell LysateLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/16498_ab32072-225518-WBGR16261511.jpg)
All lanes : Anti-c-Myc antibody [Y69] (ab32072) at 1/1000 dilution (unpurified)Lane 1 : Raji (Human Burkitt's lymphoma cell line) Whole Cell LysateLane 2 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell LysateLane 3 : THP1 (Human acute monocytic leukemia cell line) Whole Cell LysateLane 4 : A20 (Mouse B lymphoma cell line) Whole Cell LysateLane 5 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell LysateLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human diffuse large B cell lymphoma tissue labelling c-Myc with purified ab32072 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of the colon tissue labelling c-Myc with purified ab32072 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skin carcinoma tissue labelling c-Myc with unpurified ab32072 at 1/50.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarinoma of colon tissue labelling c-Myc with unpurified ab32072.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung adenocarinoma tissue labelling c-Myc with unpurified ab32072.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric adenocarcinoma tissue labelling c-Myc with unpurified ab32072.
Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human urinary bladder transitional carcinoma tissue labelling c-Myc with unpurified ab32072.
![c-Myc was immunoprecipitated using 0.5mg Jurkat 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 ab32072.Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.Band: 57kDa; c-Myc [Y69]](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/16506_ab32072-197157-AbBeadsIPab3207212m.jpg)
c-Myc was immunoprecipitated using 0.5mg Jurkat 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 ab32072.Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.Band: 57kDa; c-Myc [Y69]
Equilibrium disassociation constant (KD)Learn more about KD Click here to learn more about KD
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