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Western Blot (WB) (Figure 1. Western blot analysis of Cyclin D3 using anti-Cyclin D3 antibody (MBS177055). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. lane 1: recombinant human Cyclin D3 protein 0.5ng. 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 D3 antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for Cyclin D3 at approximately 25KD. The expected band size for Cyclin D3 is at 25KD. )

Rabbit anti-Human, Rat Cyclin D3 Polyclonal Antibody | anti-CCND3 antibody

Anti-Cyclin D3 Antibody

Reactivity
Human, Rat
Applications
Western Blot
Purity
Immunogen affinity purified.
Synonyms
Cyclin D3; Polyclonal Antibody; Anti-Cyclin D3 Antibody; G1/S-specific cyclin-D3; cyclin D3; CCND 3 antibody; CCND3 antibody; CCND3_HUMAN antibody; CyclinD3 antibody; D3 type cyclin antibody; G1 S specific cyclin D3 antibody; G1/S specific cyclin D3 antibody; G1/S-specific cyclin-D3 antibody; anti-CCND3 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Rat
Clonality
Polyclonal
Isotype
IgG
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized
Sequence Length
292
Applicable Applications for anti-CCND3 antibody
Western Blot (WB)
Immunogen
E Coli-derived human Cyclin D3 recombinant protein (Position: Q136-L292). Human Cyclin D3 shares 94% and 93% amino acid (aa) sequences identity with mouse and rat Cyclin D3, respectively.
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Preparation and Storage
At -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of Cyclin D3 using anti-Cyclin D3 antibody (MBS177055). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. lane 1: recombinant human Cyclin D3 protein 0.5ng. 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 D3 antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for Cyclin D3 at approximately 25KD. The expected band size for Cyclin D3 is at 25KD. )

Western Blot (WB) (Figure 1. Western blot analysis of Cyclin D3 using anti-Cyclin D3 antibody (MBS177055). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. lane 1: recombinant human Cyclin D3 protein 0.5ng. 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 D3 antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for Cyclin D3 at approximately 25KD. The expected band size for Cyclin D3 is at 25KD. )

Western Blot (WB)

(Figure 2. Western blot analysis of Cyclin D4 using anti-Cyclin D4 antibody (MBS177055). 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: rat thymus tissue lysate, lane 3: rat lung tissue lysate, lane 4: rat ovary tissue lysate, lane 5: JURKAT whole cell lysate, lane 6: A549 whole cell lysate, lane 7: MCF-7 whole cell lysate, lane 8: HELA 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 D4 antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for Cyclin D4 at approximately 33KD. The expected band size for Cyclin D4 is at 33KD. )

Western Blot (WB) (Figure 2. Western blot analysis of Cyclin D4 using anti-Cyclin D4 antibody (MBS177055). 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: rat thymus tissue lysate, lane 3: rat lung tissue lysate, lane 4: rat ovary tissue lysate, lane 5: JURKAT whole cell lysate, lane 6: A549 whole cell lysate, lane 7: MCF-7 whole cell lysate, lane 8: HELA 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 D4 antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for Cyclin D4 at approximately 33KD. The expected band size for Cyclin D4 is at 33KD. )

Flow Cytometry (FC/FACS)

(Figure 3. Flow Cytometry analysis of K562 cells using anti-Cyclin D3 antibody (MBS177055).Overlay histogram showing K562 cells stained with MBS177055 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Cyclin D3 Antibody (MBS177055,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 3. Flow Cytometry analysis of K562 cells using anti-Cyclin D3 antibody (MBS177055).Overlay histogram showing K562 cells stained with MBS177055 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Cyclin D3 Antibody (MBS177055,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS)

(Figure 4. Flow Cytometry analysis of THP-1 cells using anti-Cyclin D3 antibody (MBS177055).Overlay histogram showing THP-1 cells stained with MBS177055 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Cyclin D3 Antibody (MBS177055,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 4. Flow Cytometry analysis of THP-1 cells using anti-Cyclin D3 antibody (MBS177055).Overlay histogram showing THP-1 cells stained with MBS177055 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Cyclin D3 Antibody (MBS177055,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Related Product Information for anti-CCND3 antibody
Description: Rabbit IgG polyclonal antibody for G1/S-specific cyclin-D3(CCND3) detection. Tested with WB in Human, Rat.
Background: CCND3, also called Cyclin D3, is a protein that in humans is encoded by the CCND3 gene. It is mapped to 6p21.1. 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 CDK4 or CDK6, whose activtiy is required for cell cycle G1/S transition. In addition, overexpression of CCND3 upregulated the translational activity in HeLa cells in a dose-dependent manner.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
896
UniProt Accession #
Molecular Weight
9,736 Da
NCBI Official Full Name
G1/S-specific cyclin-D3
NCBI Official Synonym Full Names
cyclin D3
NCBI Official Symbol
CCND3
NCBI Protein Information
G1/S-specific cyclin-D3; D3-type cyclin; G1/S-specific cyclin D3
UniProt Protein Name
G1/S-specific cyclin-D3
Protein Family
UniProt Gene Name
CCND3
UniProt Entry Name
CCND3_HUMAN

NCBI Description

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 CDK4 or CDK6, whose activtiy is required for cell cycle G1/S transition. This protein has been shown to interact with and be involved in the phosphorylation of tumor suppressor protein Rb. The CDK4 activity associated with this cyclin was reported to be necessary for cell cycle progression through G2 phase into mitosis after UV radiation. Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Oct 2008]

Uniprot Description

CCND3: Regulatory component of the cyclin D3-CDK4 (DC) complex that phosphorylates and inhibits members of the retinoblastoma (RB) protein family including RB1 and regulates the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complex and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also substrate for SMAD3, phosphorylating SMAD3 in a cell-cycle-dependent manner and repressing its transcriptional activity. Component of the ternary complex, cyclin D3/CDK4/p27Kip1, required for nuclear translocation and activity of the cyclin D-CDK4 complex. Interacts with the CDK4 and CDK6 protein kinases to form a serine/threonine kinase holoenzyme complex. The cyclin subunit imparts substrate specificity to the complex. Interacts with ATF5. Interacts with EIF3K. Component of the ternary complex cyclin D/CDK4/p27Kip1 required for nuclear translocation and modulation of CDK4-mediated kinase activity. Can form similar complexes with either p21Cip1 or CDKN2A. Belongs to the cyclin family. Cyclin D subfamily. 2 isoforms of the human protein are produced by alternative splicing.

Protein type: Cell cycle regulation; Nuclear receptor co-regulator; Activator

Chromosomal Location of Human Ortholog: 6p21

Cellular Component: membrane; cytoplasm; cyclin-dependent protein kinase holoenzyme complex; nucleus

Molecular Function: protein binding; cyclin-dependent protein kinase activity; protein kinase binding

Biological Process: T cell proliferation; positive regulation of cyclin-dependent protein kinase activity; cell division; positive regulation of protein amino acid phosphorylation; cell cycle; signal transduction

Research Articles on CCND3

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Product Notes

The CCND3 ccnd3 (Catalog #AAA177055) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-Cyclin D3 Antibody reacts with Human, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's Cyclin D3 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). Researchers should empirically determine the suitability of the CCND3 ccnd3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Cyclin D3, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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