重组Anti-Cullin 2/CUL-2抗体[EPR3104(2)]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cullin 2/CUL-2 antibody [EPR3104(2)] (AB166917)

Intracellular Flow Cytometry analysis of 293T (Human embryonic kidney epithelial cell) cells labeling Cullin 2/CUL-2 (red) with ab166917 at a 1/100 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

• IP

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Immunoprecipitation - Anti-Cullin 2/CUL-2 antibody [EPR3104(2)] (AB166917)

Western blot analysis of an immunoprecipitation pellet from Raji cell lysate labeling Cullin 2/CUL-2 with ab166917 which was used at 1/10 dilution for immunoprecipitation.
Secondary antibody used was an HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

All lanes:

Immunoprecipitation - Anti-Cullin 2/CUL-2 antibody [EPR3104(2)] (ab166917)

Predicted band size: 87 kDa

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• WB

Supplier Data

Western blot - Anti-Cullin 2/CUL-2 antibody [EPR3104(2)] (AB166917)

Blocking and Diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Cullin 2/CUL-2 antibody [EPR3104(2)] (ab166917) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Mouse spleen lysate at 10 µg

Lane 5:

Rat brain lysate at 10 µg

Lane 6:

Rat heart lysate at 10 µg

Lane 7:

Rat kidney lysate at 10 µg

Lane 8:

Rat spleen lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 87 kDa

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Exposure time: 3min

• WB

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Western blot - Anti-Cullin 2/CUL-2 antibody [EPR3104(2)] (AB166917)

All lanes:

Western blot - Anti-Cullin 2/CUL-2 antibody [EPR3104(2)] (ab166917) at 1/1000 dilution

Lane 1:

PC3 cell lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Lane 4:

Raji cell lysate at 10 µg

Predicted band size: 87 kDa

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• WB

CiteAb

Western blot - Anti-Cullin 2/CUL-2 antibody [EPR3104(2)] (AB166917)

Cullin 2/CUL-2 western blot using anti-Cullin 2/CUL-2 antibody [EPR3104(2)] ab166917. Publication image and figure legend from Chen, K., Zeng, J., et al., 2016, J Cell Sci, PubMed 26743088.

ab166917 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab166917 please see the product overview.

p62 interacts with the VHL E3 ubiquitin ligase complex. (A) p62 interacts with CUL2. Co-transfection of Myc–CUL2 into HEK293T cells was performed, together with Flag–VHL or Flag–p62. At 48 h after transfection, the whole-cell lysates were extracted for co-immunoprecipitation with an anti-Flag antibody, followed by probing for Myc. IB, immunoblot; IP, immunoprecipitation. (B) HEK293T cells were co-transfected with HA–p62 together with Flag–VHL, Flag–CUL2, Flag–elonginB, Flag–elonginC, Flag–HIF1α or psi-Flag. At 48 h after transfection, the whole-cell lysates were extracted for co-immunoprecipitation with an antibody against Flag, followed by probing with an anti-HA antibody. (C) p62 interacts with VHL and CUL2 in vivo. The HEK293T cell lysates were immunoprecipitated (IP) with a control antibody (rabbit IgG) or an anti-­p62 antibody, and analyzed by immunoblot (IB) with antibodies against VHL, CUL2 or HIF1α. (D) Domain structure and deletion constructs of VHL (left) and p62 (right). Numbers refer to amino acid residues. (E) Mapping of the p62 ­binding region of VHL. HEK293T cells were transiently transfected with HA–p62 along with various Flag-­tagged VHL deletion mutants, as indicated. The cell lysates were immunoprecipitated with an anti-­Flag antibody and immunoblotted with an anti-­HA antibody. (F) Mapping of the VHL- and CUL2-binding region of p62. HEK293T cells were transiently transfected with HA–VHL and Myc–CUL2 along with various Flag-tagged p62 deletion mutants, as indicated. The cell lysates were immunoprecipitated with anti-­Flag antibody and immunoblotted with anti-­HA and anti-Myc antibodies.

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