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Product Info Summary
| SKU: | A00543-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-Cyclin E1/CCNE1 Antibody Picoband®
SKU/Catalog Number
A00543-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Cyclin E1/CCNE1 Antibody Picoband® catalog # A00543-2. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Cyclin E1/CCNE1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00543-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Cyclin E1/CCNE1 recombinant protein (Position: R3-A386).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00543-2 is reactive to CCNE1 in Human, Mouse, Rat
Observed Molecular Weight
47 kDa
Calculated molecular weight
47.1 kDa
Background of CCNE1
G1/S-specific cyclin-E1 is a protein that in humans is encoded by the CCNE1 gene. It is mapped to 19q12. The protein encoded by this gene belongs to the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance through the cell cycle. Cyclins function as regulators of CDK kinases. Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. This cyclin forms a complex with and functions as a regulatory subunit of CDK2, whose activity is required for cell cycle G1/S transition. This protein accumulates at the G1-S phase boundary and is degraded as cells progress through S phase. Overexpression of this gene has been observed in many tumors, which results in chromosome instability, and thus may contribute to tumorigenesis. This protein was found to associate with, and be involved in, the phosphorylation of NPAT protein (nuclear protein mapped to the ATM locus), which participates in cell-cycle regulated histone gene expression and plays a critical role in promoting cell-cycle progression in the absence of pRB.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00543-2 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.25μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human Hela whole cell, human HepG2 whole cell, human K562 whole cell, human U2OS whole cell, human A549 whole cell, human PANC-1 whole cell, human CACO-2 whole cell,
IHC: human placenta tissue, human placenta tissue, human lung cancer tissue, mouse testis tissue, rat testis tissue
FCM: SiHa cell
Validation Images & Assay Conditions
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Western blot analysis of CCNE1 using anti-CCNE1 antibody (A00543-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human U2OS whole cell lysates,
Lane 5: human A549 whole cell lysates,
Lane 6: human PANC-1 whole cell lysates,
Lane 7: human CACO-2 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CCNE1 antigen affinity purified polyclonal antibody (Catalog # A00543-2) at 0.25 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CCNE1 at approximately 47KD. The expected band size for CCNE1 is at 47KD.
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IHC analysis of CCNE1 using anti-CCNE1 antibody (A00543-2).
CCNE1 was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CCNE1 Antibody (A00543-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
IHC analysis of CCNE1 using anti-CCNE1 antibody (A00543-2).
CCNE1 was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CCNE1 Antibody (A00543-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
IHC analysis of CCNE1 using anti-CCNE1 antibody (A00543-2).
CCNE1 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CCNE1 Antibody (A00543-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
IHC analysis of CCNE1 using anti-CCNE1 antibody (A00543-2).
CCNE1 was detected in paraffin-embedded section of mouse testis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CCNE1 Antibody (A00543-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
IHC analysis of CCNE1 using anti-CCNE1 antibody (A00543-2).
CCNE1 was detected in paraffin-embedded section of rat testis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CCNE1 Antibody (A00543-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Flow Cytometry analysis of SiHa cells using anti-CCNE1 antibody (A00543-2).
Overlay histogram showing SiHa cells stained with A00543-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CCNE1 Antibody (A00543-2, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-Cyclin E1/CCNE1 Antibody Picoband® (A00543-2)
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