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Product Info Summary
| SKU: | PA1245-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, WB |
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Product info
Product Name
Anti-Cyclin D1/CCND1 Antibody Picoband®
SKU/Catalog Number
PA1245-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Cyclin D1/CCND1 Antibody catalog # PA1245-2. Tested in Flow Cytometry, WB applications. This antibody reacts with Human, 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 D1/CCND1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1245-2)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the N-terminus of human Cyclin D1, different from the related mouse and rat sequences by two amino acids.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PA1245-2 is reactive to CCND1 in Human, Rat
Observed Molecular Weight
33 kDa
Calculated molecular weight
33.7 kDa
Background of CCND1
Cyclin D1, also known as CCND1, is a human gene. The protein encoded by this gene belongs to the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance throughout the cell cycle. Cyclin D1 encodes the regulatory subunit of a holoenzyme that phosphorylates and inactivates the retinoblastoma protein and promotes progression through the G1-S phase of the cell cycle. Amplification or overexpression of cyclin D1 plays pivotal roles in the development of a subset of human cancers including parathyroid adenoma, breast cancer, colon cancer, lymphoma, melanoma, and prostate cancer. The cyclin D1 gene is overexpressed in human breast cancers and is required for oncogene-induced tumorigenesis. Brisken et al. (2003) found that prolactin (PRL; 176760) induced IGF2 (147470) mRNA and IGF2 induced cyclin D1 protein expression in mouse mammary epithelial cultures. And they also concluded that IGF2 is a mediator of prolactin-induced alveologenesis and that prolactin, IGF2, and cyclin D1 are components of a developmental pathway in mammary gland.
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
PA1245-2 is guaranteed for 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, 0.1-0.5μg/ml, Human, Rat
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: Rat Testis Tissue, Human Placenta Tissue, Rat Brain Tissue, MCF-7 Whole Cell, COLO320 Whole Cell, SW620 Whole Cell, MM231 Whole Cell
FCM: A431 cell
Validation Images & Assay Conditions
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Western blot analysis of Cyclin D1 using anti-Cyclin D1 antibody (PA1245-2).
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: Rat Testis Tissue Lysate,
Lane 2: Human Placenta Tissue Lysate,
Lane 3: Rat Brain Tissue Lysate,
Lane 4: MCF-7 Whole Cell Lysate,
Lane 5: COLO320 Whole Cell Lysate,
Lane 6: SW620 Whole Cell Lysate,
Lane 7: MM231 Whole Cell Lysate.
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-Cyclin D1 antigen affinity purified polyclonal antibody (Catalog # PA1245-2) at 0.5 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 1:10000 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 Cyclin D1 at approximately 33KD. The expected band size for Cyclin D1 is at 33KD.
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Flow Cytometry analysis of A431 cells using anti-Cyclin D1 antibody (PA1245-2).
Overlay histogram showing A431 cells stained with PA1245-2 (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-Cyclin D1 Antibody (PA1245-2, 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-Cyclin D1/CCND1 Antibody Picoband® (PA1245-2)
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