Anti-Cyclin E1 antibody [EPR25394-23] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E1 antibody [EPR25394-23] - BSA and Azide free (AB317437)

This data was developed using ab317436, 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 Cyclin E1 with ab317436 at 1/500 (1.004 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), and decreased staining in cells during mitosis (indicated by arrowhead). 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.

• WB

Supplier Data

Western blot - Anti-Cyclin E1 antibody [EPR25394-23] - BSA and Azide free (AB317437)

This data was developed using ab317436, the same antibody clone in a different buffer formulation.

The high-sensitivity ECL substrate allows for the detection of proteins in the mid-femtogram range.

Low expression : heart, kidney (PMID : 9840943, PMID : 11687642).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

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

All lanes:

Western blot - Anti-Cyclin E1 antibody [EPR25394-23] (<a href='/ko/products/primary-antibodies/cyclin-e1-antibody-epr25394-23-ab317436'>ab317436</a>) at 1/1000 dilution

Lane 1:

Rat testis tissue lysate at 20 µg

Lane 2:

Rat spleen tissue lysate at 20 µg

Lane 3:

Rat heart tissue lysate at 20 µg

Lane 4:

Rat kidney tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 50 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Cyclin E1 antibody [EPR25394-23] - BSA and Azide free (AB317437)

This data was developed using ab317436, the same antibody clone in a different buffer formulation.

The high-sensitivity ECL substrate allows for the detection of proteins in the mid-femtogram range.

Low expression : lung, heart, kidney (PMID : 9840943, PMID : 11687642).

The identity of the higher molecular weight band at approximately 100 kDa may represent multi-ubiquitylated Cyclin E1 (PMID : 23109421). This has not been confirmed with additional tests.

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

Exposure time : Lane 1 : 70 seconds, lanes 2-6 : 180 seconds

All lanes:

Western blot - Anti-Cyclin E1 antibody [EPR25394-23] (<a href='/ko/products/primary-antibodies/cyclin-e1-antibody-epr25394-23-ab317436'>ab317436</a>) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse placenta tissue lysate at 20 µg

Lane 3:

Mouse testis tissue lysate at 20 µg

Lane 4:

Mouse lung tissue lysate at 20 µg

Lane 5:

Mouse kidney tissue lysate at 20 µg

Lane 6:

Mouse heart tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 50 kDa,100 kDa,36 kDa

true

• WB

Supplier Data

Western blot - Anti-Cyclin E1 antibody [EPR25394-23] - BSA and Azide free (AB317437)

This data was developed using ab317436, the same antibody clone in a different buffer formulation.

The expression of Cyclin E1 is upregulated in response to aphidicolin treatment (PMID : 29995582).

The identity of the higher molecular weight band at approximately 100 kDa may represent multi-ubiquitylated Cyclin E1 (PMID : 23109421). This has not been confirmed with additional tests.

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

Exposure time : Lanes 1-2 : 180 seconds, lanes 3-4 : 103 seconds

All lanes:

Western blot - Anti-Cyclin E1 antibody [EPR25394-23] (<a href='/ko/products/primary-antibodies/cyclin-e1-antibody-epr25394-23-ab317436'>ab317436</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

NIH/3T3 transfected with siRNA specifically targeting Cyclin E1 whole cell lysate at 20 µg

Lane 3:

Untreated NIH/3T3 whole cell lysate at 20 µg

Lane 4:

NIH/3T3 treated with 10 µg/ml aphidicolin for 24 hours whole cell lysate

Secondary

All lanes:

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

Observed band size: 50 kDa,100 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Cyclin E1 antibody [EPR25394-23] - BSA and Azide free (AB317437)

This data was developed using ab317436, the same antibody clone in a different buffer formulation.

This antibody does not cross-react with mouse Cyclin E2.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Cyclin E1 antibody [EPR25394-23] (<a href='/ko/products/primary-antibodies/cyclin-e1-antibody-epr25394-23-ab317436'>ab317436</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a mouse Cyclin E1 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a mouse Cyclin E2 expression vector containing a His-tag, whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 50 kDa,124 kDa

false

Exposure time: 10s