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Cyclin D3 Recombinant Mouse mAb

Cell line: HeLa Fixative: 4% Paraformaldehyde Permeabilization: 0.1% TritonX-100 Primary ab dilution: 1:50 Primary incubation condition: 4°C overnight Secondary ab: Goat Anti-Mouse IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for AP94609
Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:1000 Primary ab incubation condition: room temperature 2h Secondary ab: Goat Anti-Mouse IgG H&L (HRP) Lysate: BRL Protein loading quantity: 20 µg Exposure time: 10 s Predicted MW: 33 kDa Observed MW: 33 kDa
Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:1000 Primary ab incubation condition: room temperature 2h Secondary ab: Goat Anti-Mouse IgG H&L (HRP) Lysate: HeLa, Daudi Protein loading quantity: 20 µg Exposure time: 10 s Predicted MW: 33 kDa Observed MW: 33 kDa

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IF, ICC
Host	Rabbit
Clonality	Recombinant
Physical State	Liquid
Isotype	IgG1, Kappa
Purity	affinity purified by Protein G
Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
SUBCELLULAR LOCATION	Nucleus. Cytoplasm. Membrane. Note=Cyclin D-CDK4 complexes accumulate at the nuclear membrane and are then translocated to the nucleus through interaction with KIP/CIP family members.
SIMILARITY	Belongs to the cyclin family. Cyclin D subfamily. Contains 1 cyclin N-terminal domain.
SUBUNIT	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/CDKN1B required for nuclear translocation and modulation of CDK4-mediated kinase activity. Can form similar complexes with either CDKN1A or CDKN2A.
Post-translational modifications	Polyubiquitinated by the SCF(FBXL2) complex, leading to proteasomal degradation.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	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].

Additional Information
Target/Specificity
Dilution	WB=1:500-1:1000,ICC/IF=1:50
Format	0.01M TBS(pH7.4) with 1% BSA, 0.09% (W/V) sodium azide and 50% Glyce
Storage	Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

Research Areas

For Research Use Only. Not For Use In Diagnostic Procedures.

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol