Cyclin T1 Recombinant Rabbit Monoclonal Antibody [PSH09-66]
cat.: HA723131
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, FC, IP, ChIP
Clonality: Monoclonal
Clone number: PSH09-66
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 81 kDa
Isotype: IgG
Immunogen: Recombinant protein within human Cyclin T1 aa 251-530.
Positive control: Jurkat cell lysate, K-562 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, NIH/3T3 cell lysate, C6 cell lysate, mouse liver tissue, rat liver tissue, NIH/3T3, C6.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC
  IP
  ChIP
1:2,000
1:2,000
1:100
1:1,000
1-2μg/sample
Use 2~5 μg for 25 μg of chromatin.
Uniprot #: SwissProt: O60563 Human | Q9QWV9 Mouse
Entrez Gene: 315291 Rat
Alternative names: CCN T1 CCNT CCNT 1 Ccnt1 CCNT1_HUMAN CDK9 associated C type protein Cyc T1 Cyclin C related protein Cyclin T cyclin T1 Cyclin T1b Cyclin-T Cyclin-T1 CYCT 1 CycT1 HIVE1 Human immunodeficiency virus 1 expression Human immunodeficiency virus type 1 (HIV 1) expression (elevated) 1 pTEFb subunit Subunit of positive elongation transcription factor b
Images
HA723131_1.jpg Fig1: Western blot analysis of Cyclin T1 on different lysates with Rabbit anti-Cyclin T1 antibody (HA723131) at 1/2,000 dilution.

Lane 1: Jurkat cell lysate (20 µg/Lane)
Lane 2: K-562 cell lysate (20 µg/Lane)
Lane 3: Mouse brain tissue lysate (40 µg/Lane)
Lane 4: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 81 kDa
Observed band size: 75 kDa

Exposure time: 25 seconds; ECL: K1802;

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 (HA723131) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA723131_2.jpg Fig2: Western blot analysis of Cyclin T1 on different lysates with Rabbit anti-Cyclin T1 antibody (HA723131) at 1/2,000 dilution.

Lane 1: NIH/3T3 cell lysate
Lane 2: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 81 kDa
Observed band size: 75 kDa

Exposure time: 1 minute; 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 (HA723131) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA723131_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Cyclin T1 antibody (HA723131) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723131) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723131_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Cyclin T1 antibody (HA723131) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723131) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723131_5.jpg Fig5: Immunocytochemistry analysis of NIH/3T3 cells labeling Cyclin T1 with Rabbit anti-Cyclin T1 antibody (HA723131) 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-Cyclin T1 antibody (HA723131) 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.
HA723131_6.jpg Fig6: Immunocytochemistry analysis of C6 cells labeling Cyclin T1 with Rabbit anti-Cyclin T1 antibody (HA723131) 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-Cyclin T1 antibody (HA723131) 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.
HA723131_7.jpg Fig7: Flow cytometric analysis of NIH/3T3 cells labeling Cyclin T1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723131, 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).
HA723131_8.jpg Fig8: Cyclin T1 was immunoprecipitated from 0.2 mg Jurkat cell lysate with HA723131 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723131 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: Jurkat cell lysate (input)
Lane 2: HA723131 IP in Jurkat cell lysate
Lane 3: Rabbit IgG instead of HA723131 in Jurkat cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 37 seconds; ECL: K1802
HA723131_9.jpg Fig9: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Cyclin T1 (HA723131) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
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