Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Cullin 4B/CUL-4B with ab317014 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling Cullin 4B/CUL-4B with ab317014 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human breast carcinoma.
The section was incubated with ab317014 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Cullin 4B/CUL-4B with ab317014 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human testis.
The section was incubated with ab317014 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Cullin 4B/CUL-4B with ab317014 at 1/50 (9.8 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing nuclear staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

Cullin 4B/CUL-4B was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab317014 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317014 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab317014 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317014 in HeLa whole cell lysate

All lanes:

Immunoprecipitation - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] (<a href='/en-us/products/primary-antibodies/cullin-4b-cul-4b-antibody-epr26157-39-ab317014'>ab317014</a>) at 1/30 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 50s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Cullin 4B/CUL-4B with ab317014 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse testis (PMID : 26832838).
The section was incubated with ab317014 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Cullin 4B/CUL-4B with ab317014 at 1/50 (9.8 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing nuclear staining in NIH/3T3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labeling Cullin 4B/CUL-4B with ab317014 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse breast cancer.
The section was incubated with ab317014 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Cullin 4B/CUL-4B with ab317014 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat testis.
The section was incubated with ab317014 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] (<a href='/en-us/products/primary-antibodies/cullin-4b-cul-4b-antibody-epr26157-39-ab317014'>ab317014</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting Cullin 4B whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 104 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

The identity of the lower MW band at approximately 37 kDa is unknown.

All lanes:

Western blot - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] (<a href='/en-us/products/primary-antibodies/cullin-4b-cul-4b-antibody-epr26157-39-ab317014'>ab317014</a>) at 1/1000 dilution

Lane 1:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 30 µg with 5% NFDM/TBST

Lane 2:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 30 µg with 5% NFDM/TBST

Lane 3:

Mouse testis tissue lysate at 30 µg with 5% NFDM/TBST

Lane 4:

Mouse liver tissue lysate at 30 µg with 5% NFDM/TBST

Lane 5:

Rat testis tissue lysate at 30 µg with 5% NFDM/TBST

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG (Merck DC03L) at 1/2000 dilution

Observed band size: 104 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] - BSA and Azide free (AB317015)

This data was developed using ab317014, the same antibody clone in a different buffer formulation.

The lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-Cullin 4B/CUL-4B antibody [EPR26157-39] (<a href='/en-us/products/primary-antibodies/cullin-4b-cul-4b-antibody-epr26157-39-ab317014'>ab317014</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 104 kDa

false

Exposure time: 180s