Anti-Cullin 1/CUL-1 抗体 [EPR3103Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

Unpurified ab75817 showing positive staining in human Ovarian carcinoma tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Cullin 1/CUL-1 with purified ab75817 at 1/20 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr^®488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

Immunohistochemical staining of paraffin embedded human lung carcinoma with purified ab75817 at a working dilution of 1/50. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

Unpurified ab75817 showing positive staining in Normal human tonsil tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

Immunofluorescence staining of HeLa cells with purified ab75817 at a working dilution of 1/1000, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab75817 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

Unpurified ab75817 at 1/100 dilution staining Cullin 1/CUL-1 in human cervical carcinoma by Immunohistochemistry, Paraffin-embedded tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

Unpurified ab75817 showing positive staining in human Urinary bladder transitional carcinoma tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab75817 at a working dilution of 1/50. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IP

Unknown

Immunoprecipitation - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

ab75817 (purified) at 1/120 immunoprecipitating Cullin 1 in 10 μg MCF7 whole cell lysate (Lanes 1 and 2, observed at 90 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution. Blocking buffer and concentration : 5% NFDM/TBST Dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

Predicted band size: 90 kDa

false

• WB

Lab

Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

MCF7 cell lysate at 10 µg

Lane 3:

A549 cell lysate at 10 µg

Lane 4:

MDA-MB-435 cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 47 kDa,90 kDa

Observed band size: 49 kDa,90 kDa

false

• WB

Lab

Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817) at 1/1000 dilution

Lane 1:

PC-12 cell lysate at 10 µg

Lane 2:

NIH/3T3 cell lysate at 10 µg

Lane 3:

mouse brain cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 90 kDa

Observed band size: 90 kDa

false

• WB

Unknown

Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (AB75817)

All lanes:

Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817) at 1/5000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

MCF7 cell lysate at 10 µg

Lane 3:

A549 cell lysate at 10 µg

Lane 4:

PC12 cell lysate at 10 µg

Secondary

All lanes:

goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 90 kDa

Observed band size: 90 kDa

false