Anti-Mad2L1 antibody

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mad2L1 antibody (AB97777)

Immunohistochemical analysis of paraffin-embedded human a549 xenograft tissue stained for MAD2L1 protein at cytoplasm and nucleus with ab97777 at a 1/250 dilution.

Antigen retrieval : EDTA based buffer, (pH 8.0), 15min

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Mad2L1 antibody (AB97777)

Immunofluorescence analysis of Mad2L1 in Human osteosarcoma cell line U2OS, using ab97777 at 1/1000 dilution.
1 : Mad2L1
2 : ACA
3 : merged with DAPI
Cells were fixed with 4% PFA/PBS for 15min, RT and blocked with 2.5% BSA/PBS.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mad2L1 antibody (AB97777)

Immunohistochemical analysis of Mad2L1 in paraffin embedded DLD-1 xenograft, using ab97777 at a 1/500 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Mad2L1 antibody (AB97777)

Immunoprecipitation of MAD2L1 protein from 293T whole cell extracts using 5 µg of ab97777. Western blot analysis was performed using ab97777.

All lanes:

Immunoprecipitation - Anti-Mad2L1 antibody (ab97777) at 5 µg

Lane 1:

293T whole cell extracts (Input)

Lane 2:

293T whole cell extracts with control IgG (-)

Lane 3:

293T whole cell extracts (+)

Secondary

All lanes:

Anti-Rabbit IgG

Predicted band size: 24 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mad2L1 antibody (AB97777)

Immunohistochemical analysis of paraffin-embedded rat fore brain tissue stained for MAD2L1 protein at nucleus with ab97777 at a 1/500 dilution.

Antigen retrieval : EDTA based buffer, (pH 8.0), 15min

• WB

Supplier Data

Western blot - Anti-Mad2L1 antibody (AB97777)

Samples were separated by 12% SDS-PAGE.

All lanes:

Western blot - Anti-Mad2L1 antibody (ab97777) at 1/1000 dilution

Lane 1:

293T whole cell extracts at 30 µg

Lane 2:

MAD2L1 shRNA transfected 293T whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 24 kDa

false

• WB

Supplier Data

Western blot - Anti-Mad2L1 antibody (AB97777)

Samples were separated by 12% SDS-PAGE.

All lanes:

Western blot - Anti-Mad2L1 antibody (ab97777) at 1/1000 dilution

Lane 1:

Jurkat whole cell extracts at 30 µg

Lane 2:

Raji whole cell extracts at 30 µg

Lane 3:

NCI-H929 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 24 kDa

false

• WB

Unknown

Western blot - Anti-Mad2L1 antibody (AB97777)

All lanes:

Western blot - Anti-Mad2L1 antibody (ab97777) at 1/1000 dilution

Lane 1:

HepG2 whole cell lysate at 30 µg

Lane 2:

MOLT4 whole cell lysate at 30 µg

Lane 3:

Raji whole cell lysate at 30 µg

Predicted band size: 24 kDa

false

• WB

CiteAb

Western blot - Anti-Mad2L1 antibody (AB97777)

Western Blotting using Anti-Mad2L1 antibodyAnti-Mad2L1 antibody, ab97777. Publication image from Chen, J. et al., 2016, Nat Commun, 26743940. Legend direct from paper.

Cep57 contributes to activation of the spindle assembly checkpoint.(a,b) HeLa cells with 100 nM nocodazole treatment after co-transfection with H2B-RFP and negative control (NC)-, Cep57-, Mad1- or Mad2-siRNA for 48 h. (a) Time-lapse images of HeLa cells. The numbers indicate the time (minutes) after entry into mitosis. Arrowhead, multiple nuclei. Scale bar, 5 µm. (b) Quantification of mitotic duration of HeLa cells from a. Scatter plots show data from three independent experiments. (c) Quantification of mitotic duration of live HeLa cells with 80 nM taxol treatment after co-transfection with H2B-RFP and negative control (NC)-, Cep57-, Mad1- or Mad2-siRNA for 48 h. Scatter plots show data from three independent experiments. (d) Diagram of experimental design. The siRNA-transfected HeLa cells were synchronized by double-thymidine block, and released into nocodazole. (e–g) HeLa cells were treated as in d. (e) After 12 h nocodazole treatment, mitotic cell lysates were used to perform immunoprecipitation (IP) and western blotting assays with the indicated antibodies. WCL, whole-cell lysate. (f) Western blots of the indicated proteins in HeLa cells after the indicated nocodazole (Noc.) treatment time. (g) Quantification of the protein levels of securin and cyclin B1 from (f). The experiment was repeated three times. For b,c and g, data are mean±s.e.m. ****P<0.0001; ***P<0.001; **P<0.01; *P<0.05; NS, not significant (unpaired two-tailed Student's t-test). DIC, differential interference contrast.

false

• WB

CiteAb

Western blot - Anti-Mad2L1 antibody (AB97777)

Western Blotting using Anti-Mad2L1 antibodyAnti-Mad2L1 antibody, ab97777. Publication image from Chen, J. et al., 2016, Nat Commun, 26743940. Legend direct from paper.

Cep57 contributes to activation of the spindle assembly checkpoint.(a,b) HeLa cells with 100 nM nocodazole treatment after co-transfection with H2B-RFP and negative control (NC)-, Cep57-, Mad1- or Mad2-siRNA for 48 h. (a) Time-lapse images of HeLa cells. The numbers indicate the time (minutes) after entry into mitosis. Arrowhead, multiple nuclei. Scale bar, 5 µm. (b) Quantification of mitotic duration of HeLa cells from a. Scatter plots show data from three independent experiments. (c) Quantification of mitotic duration of live HeLa cells with 80 nM taxol treatment after co-transfection with H2B-RFP and negative control (NC)-, Cep57-, Mad1- or Mad2-siRNA for 48 h. Scatter plots show data from three independent experiments. (d) Diagram of experimental design. The siRNA-transfected HeLa cells were synchronized by double-thymidine block, and released into nocodazole. (e–g) HeLa cells were treated as in d. (e) After 12 h nocodazole treatment, mitotic cell lysates were used to perform immunoprecipitation (IP) and western blotting assays with the indicated antibodies. WCL, whole-cell lysate. (f) Western blots of the indicated proteins in HeLa cells after the indicated nocodazole (Noc.) treatment time. (g) Quantification of the protein levels of securin and cyclin B1 from (f). The experiment was repeated three times. For b,c and g, data are mean±s.e.m. ****P<0.0001; ***P<0.001; **P<0.01; *P<0.05; NS, not significant (unpaired two-tailed Student's t-test). DIC, differential interference contrast.

false