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Anti-MCL1 Antibody Y37
Also for MCL1 (NM_021960)
|A synthetic peptide corresponding to residues of human Mcl-1 was used as immunogen. The antibody does not cross-react with other Bcl-2 family members.|
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||ICC/IF: 1:100; WB: 1:1000; IHC-P: Use at an assay dependent dilution; ICC: 1:250 - 1:500; FC: Use 1?g for 106<:sup> cells; IP: 1:100
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|IgG fraction (Protein A or G Sepharose)
|Homo sapiens myeloid cell leukemia sequence 1 (BCL2-related) (MCL1), transcript variant 1|
|bcl2-L-3; BCL2L3; EAT; Mcl-1; MCL1-ES; mcl1/EAT; MCL1L; MCL1S; TM|
|The protein encoded by this gene belongs to the Bcl-2 family. Alternative splicing occurs at this locus and two transcript variants encoding distinct isoforms have been identified. The longer gene product (isoform 1) enhances cell survival by inhibiting apoptosis while the alternatively spliced shorter gene product (isoform 2) promotes apoptosis and is death-inducing. [provided by RefSeq]. |
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Western blot - MCL1 antibody [Y37]; Anti-MCL1 antibody [Y37] at 1/1000 dilution + A431 cell lysate.Predicted band size : 37 kDa.Observed band size : 37 kDa.
Immunohistochemistry (Paraffin-embedded sections) - MCL1 antibody [Y37]; Immunohistochemical analysis of MCL1 in paraffin-embedded human tonsils using 1/50 ab32087.
Immunohistochemistry (Paraffin-embedded sections) - MCL1 antibody [Y37]; Immunohistochemical analysis of MCl1 expression in paraffin-embedded cervical carcinoma using 1/50 ab32087.
Immunocytochemistry/ Immunofluorescence - Anti-MCL1 antibody [Y37]; staining MCL1 in HCT116 cells treated with wogonin , by ICC/IF. Decrease of MCL1 expression correlates with increased concentration of wogonin, as described in literature.The cells were incubated at 37°C for 2h in media containing different concentrations of (wogonin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with (1/100) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Immunocytochemistry/ Immunofluorescence - MCL1 antibody [Y37]; ICC/IF image of ab32087 stained MCF7 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 overnight at +4°C. The secondary antibody (green) was Alexa Fluor? 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.
Immunocytochemistry/ Immunofluorescence - MCL1 antibody [Y37]; ab32087 staining MCL1 in H1299 cells by Immunocytochemistry/ Immunoflourescence. Cells were PFA-fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 3% Serum for 1 hour at 24°C. The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 24°C. The secondary antibody was an Alexa Fluor? 488-conjugated Goat anti-Rabbit polyclonal, diluted 1/2000.
Flow Cytometry-MCL1 antibody [Y37](ab32087); Overlay histogram showing A431 cells stained with ab32087 (red line). The cells were fixed with 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 for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in A431 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.