MA5-14336

antibody from Invitrogen Antibodies

Targeting: CCNE1 CCNE

Antibody data

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Validation data

Product number: MA5-14336 - Provider product page
Provider: Invitrogen Antibodies
Product name: Cyclin E Monoclonal Antibody (HE12)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-14336 targets Cyclin E in FACS, IF, IHC (P), and WB applications and shows reactivity with Human and Mouse samples.
Antibody clone number: HE12
Concentration: 0.2 mg/mL

CCNE1 protein structure - MA5-14336 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Cyclin E using Cyclin E Monoclonal Antibody (Product # MA5-14336) on LS174T Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: A549 cells were lysed 72 hours after transfection. Cells were transfected with Transfection Reagent alone (Lane 1), 100nM ON-TARGETplus siCONTROL Non-Targeting Pool (Lane 2), or 100nM ONTARGETplus CCNE1 siRNA (Lane 3). Western blot data for Cyclophilin B is included as a control for equal protein loading.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Cyclin E using Cyclin E Monoclonal Antibody (Product # MA5-14336) on SKBR3 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Cyclin E using Cyclin E Monoclonal Antibody (Product # MA5-14336) on LS174T Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: A549 cells were lysed 72 hours after transfection. Cells were transfected with Transfection Reagent alone (Lane 1), 100nM ON-TARGETplus siCONTROL Non-Targeting Pool (Lane 2), or 100nM ONTARGETplus CCNE1 siRNA (Lane 3). Western blot data for Cyclophilin B is included as a control for equal protein loading.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of CCNE1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR745911_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of CCNE1 was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa CAS9 (Lane 2) and HeLa CCNE1 KO (Lane 3) modified whole cell extracts. The samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-Cyclin E Monoclonal Antibody (HE12) using 1 µg/mL dilution (Product # MA5-14336) and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to CCNE1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Cyclin E in C2C12 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant, adding 90% methanol and incubated for 10 minutes at room temperature. Follwing penetration, cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Cyclin E monoclonal antibody (Product # MA5-14336) at a dilution of 2 µg/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Cyclin E in Jurkat cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant, adding 90% methanol and incubated for 10 minutes at room temperature. Follwing penetration, cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Cyclin E monoclonal antibody (Product # MA5-14336) at a dilution of 2 µg/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Cyclin E in MCF-7 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant, adding 90% methanol and incubated for 10 minutes at room temperature. Follwing penetration, cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Cyclin E monoclonal antibody (Product # MA5-14336) at a dilution of 1 µg/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.