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Product Info Summary
| SKU: | A00257-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, WB |
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Product info
Product Name
Anti-E2F1 Antibody Picoband®
SKU/Catalog Number
A00257-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-E2F1 Antibody Picoband® catalog # A00257-2. Tested in ELISA, Flow Cytometry, 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
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 freezing and thawing.
Cite This Product
Anti-E2F1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00257-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human E2F1 recombinant protein (Position: K89-A328).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00257-2 is reactive to E2F1 in Human, Mouse, Rat
Observed Molecular Weight
70 kDa
Calculated molecular weight
46.9 kDa
Background of E2F1
Transcription factor E2F1 is a protein that in humans is encoded by the E2F1 gene. The protein encoded by this gene is a member of the E2F family of transcription factors. The E2F family of transcription factors appears to play a critical role in the transcription of certain genes required for cell cycle progression. E2F1, the first cloned member of this family, is regulated during the cell cycle at the mRNA level by changes in transcription of the E2F1 gene and at the protein level by complex formation with proteins such as the retinoblastoma gene product(pRB), cyclin A and DP1. E2F1 can override a pRB-induced G1/S block and can behave as an oncogene in certain cells. E2F1 was cloned and was found to contain seven exons. Fluorescence in situ hybridization localized E2F1 to chromosome 20q11. The E2F1 transcription factor can promote proliferation or apoptosis when activated, and is a key downstream target of the retinoblastoma tumour suppressor protein(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
A00257-2 is guaranteed for ELISA, Flow Cytometry, 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, 0.1-0.25 μg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human Hela whole cell, human A431 whole cell, human Caco-2 whole cell, rat brain tissue, rat testis tissue, mouse brain tissue, mouse testis tissue
FCM: PC-3 cell
Validation Images & Assay Conditions
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Western blot analysis of E2F1 using anti-E2F1 antibody (A00257-2).
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 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: human Caco-2 whole cell lysates,
Lane 4: rat brain tissue lysates,
Lane 5: rat testis tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse testis tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-E2F1 antigen affinity purified polyclonal antibody (Catalog # A00257-2) 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 E2F1 at approximately 70 kDa. The expected band size for E2F1 is at 47 kDa.
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Flow Cytometry analysis of PC-3 cells using anti-E2F1 antibody (A00257-2).
Overlay histogram showing PC-3 cells stained with A00257-2 (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-E2F1 Antibody (A00257-2, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/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-E2F1 Antibody Picoband® (A00257-2)
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