Cdc34 Recombinant Rabbit Monoclonal Antibody [JJ086-07]
Catalog# ET1701-10
Cdc34 Recombinant Rabbit Monoclonal Antibody [JJ086-07]
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WB
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IF-Cell
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FC
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IP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Cdc34 Recombinant Rabbit Monoclonal Antibody [JJ086-07]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human Cdc34 aa 1-236 / 236.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, FC, IP
Molecular Weight
Predicted band size: 27 kDa
Positive Control
293T cell lysate, K-562 cell lysate, Jurkat cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, RAW264.7, PC-12.
Conjugation
unconjugated
Clone Number
JJ086-07
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:100
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FC
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1:1,000
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IP
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1-2μg/sample
Target
Function
CDC34 is a gene that in humans encodes the protein Ubiquitin-conjugating enzyme E2 R1. This protein is a member of the ubiquitin-conjugating enzyme family, which catalyzes the covalent attachment of ubiquitin to other proteins. CDC34 was originally discovered by work in baker's yeast as a gene that is essential for the cell cycle. Cdc34 in yeast targets numerous substrates - notably the cyclin-dependent kinase inhibitor Sic1 for ubiquitin-mediated protein degradation. CDC34 is required for ubiquitin-mediated degradation of cell cycle G1 regulators, and for the initiation of DNA replication.
Background References
1. Dimova N.V., et al. 2012. APC/C-mediated multiple monoubiquitylation provides an alternative degradation signal for cyclin B1. Nat. Cell Biol. 14:168-176.
2. Kim C.W., et al. 2012. Ectopic over-expression of tristetraprolin in human cancer cells promotes biogenesis of let-7 by down-regulation of Lin28. Nucleic Acids Res. 40:3856-3869.
Sequence Similarity
Belongs to the ubiquitin-conjugating enzyme family.
Tissue Specificity
Expressed in testes during spermatogenesis to regulate repression of cAMP-induced transcription.
Post-translational Modification
Autoubiquitinated. Autoubiquitination is promoted by the human herpes virus 1 protein ICP0 and leads to degradation by the Ubiquitin-proteasomal pathway.; Phosphorylated by CK2. Phosphorylation of the C-terminal tail by CK2 controles the nuclear localization.
Subcellular Location
Cytoplasm, Nucleus.
Synonyms
Cdc 34 antibody
Cdc34 antibody
Cell division cycle 34 antibody
Cell division cycle 34 homolog (S. cerevisiae) antibody
Cell division cycle 34 homolog antibody
E2 CDC34 antibody
UB2R1_HUMAN antibody
UBC 3 antibody
UBC3 antibody
UBCH3 antibody
ExpandImages
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Western blot analysis of Cdc34 on different lysates with Rabbit anti-Cdc34 antibody (ET1701-10) at 1/5,000 dilution.
Lane 1: 293T cell lysate (20 µg/Lane)
Lane 2: K-562 cell lysate (20 µg/Lane)
Lane 3: Jurkat cell lysate (20 µg/Lane)
Lane 4: RAW264.7 cell lysate (20 µg/Lane)
Lane 5: PC-12 cell lysate (20 µg/Lane)
Lane 6: Mouse brain tissue lysate (40 µg/Lane)
Lane 7: Rat brain tissue lysate (40 µg/Lane)
Predicted band size: 27 kDa
Observed band size: 32 kDa
Exposure time: 8 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-10) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of RAW264.7 cells labeling Cdc34 with Rabbit anti-Cdc34 antibody (ET1701-10) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cdc34 antibody (ET1701-10) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of PC-12 cells labeling Cdc34 with Rabbit anti-Cdc34 antibody (ET1701-10) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cdc34 antibody (ET1701-10) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of RAW264.7 cells labeling Cdc34.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-10, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Flow cytometric analysis of PC-12 cells labeling Cdc34.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-10, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Cdc34 was immunoprecipitated from 0.2 mg 293T cell lysate with ET1701-10 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1701-10 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: 293T cell lysate (input)
Lane 2: ET1701-10 IP in 293T cell lysate
Lane 3: Rabbit IgG instead of ET1701-10 in 293T cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 6 seconds; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"