Cdc34 Recombinant Rabbit Monoclonal Antibody [JJ086-07]
cat.: ET1701-10
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC, IP
Clonality: Monoclonal
Clone number: JJ086-07
Form: Liquid
Storage condition: 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.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 27 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human Cdc34 aa 1-236 / 236.
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.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  FC
  IP
1:5,000
1:100
1:1,000
1-2μg/sample
Uniprot #: SwissProt: P49427 Human | Q8CFI2 Mouse
Unigene: 2427 Rat
Alternative names: Cdc 34 Cdc34 Cell division cycle 34 Cell division cycle 34 homolog (S. cerevisiae) Cell division cycle 34 homolog E2 CDC34 UB2R1_HUMAN UBC 3 UBC3 UBCH3 UBE2 R1 UBE2R1 Ubiquitin carrier protein Ubiquitin conjugating enzyme Cdc34 Ubiquitin conjugating enzyme E2 32 kDa complementing Ubiquitin protein ligase Ubiquitin protein ligase R1 Ubiquitin-conjugating enzyme E2 R1 Ubiquitin-conjugating enzyme E2-32 kDa complementing Ubiquitin-conjugating enzyme E2-CDC34 Ubiquitin-protein ligase R1
Images
ET1701-10_1.jpg Fig1: 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.
ET1701-10_2.jpg Fig2: 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.
ET1701-10_3.jpg Fig3: 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.
ET1701-10_4.jpg Fig4: 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).
ET1701-10_5.jpg Fig5: 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).
ET1701-10_6.jpg Fig6: 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
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.