Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide free (AB238940)

ab184703 staining Cyclin T1 in Jurkat (human T cell leukemia T lymphocyte). Cells were fixed with 100% Methanol. Samples were incubated with primary antibody at 1/100 dilution (6.4µg/ml). An Alexa Fluor^® 488 Goat anti-rabbit (ab150077) was used as the secondary antibody at 1/1000 dilution (2 μg/ml). ab195888, anti-alpha Tubulin [DM1A] – Microtubule Marker (Alexa Fluor^® 594) was used as counterstain antibody at 1/200 dilution (2.5 μg/ml). DAPI was used as a nuclear counterstain. Confocal image showing nuclear staining in Jurkat cell line.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184703).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide free (AB238940)

ab184703 staining Cyclin T1 in MCF7 (human breast adenocarcinoma epithelial cell). Cells were fixed with 100% Methanol. Samples were incubated with primary antibody at 1 : 100 dilution (6.4 μg/ml). An Alexa Fluor^® 488 Goat anti-rabbit (ab150077) was used as the secondary antibody at 1/1000 dilution (2 μg/ml). ab195888, anti-alpha Tubulin [DM1A] – Microtubule Marker (Alexa Fluor^® 594) was used as counterstain antibody at 1/200 dilution (2.5 μg/ml). DAPI was used as a nuclear counterstain. Confocal image showing nuclear staining in MCF7 cell line.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184703).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide free (AB238940)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Cyclin T1 with ab184703 at 1/120 dilution (red) compared with a Rabbit IgG,monoclonal [EPR17982] - Isotype Control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184703).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide free (AB238940)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Cyclin T1 with ab184703 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on hepatocytes of mouse liver is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184703).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide free (AB238940)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Cyclin T1 with ab184703 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on rat kidney is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184703).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide free (AB238940)

Cyclin T1 was immunoprecipitated from 1mg of PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate with ab184703 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab184703 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : PC-12 whole cell lysate 10μg (Input).
Lane 2 : ab184703 IP in PC-12 whole cell lysate.
Lane 3 : Rabbit IgG,monoclonal [EPR17982] - Isotpe Control (ab172730) instead of ab184703 in PC-12 whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184703).

All lanes:

Immunoprecipitation - Anti-Cyclin T1 antibody [EPR17982] (<a href='/ko/products/primary-antibodies/cyclin-t1-antibody-epr17982-ab184703'>ab184703</a>)

Predicted band size: 81 kDa

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• WB

CiteAb

Western blot - Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide free (AB238940)

Western Blotting using Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide freeAnti-Cyclin T1 antibody [EPR17982] - BSA and Azide free, ab238940. Publication image from Kress, T. R. et al., 2020, Nat Commun, 32286286. Legend direct from paper.

The juvenile heart is permissive to Myc-driven transcriptional activation.a Immunoblot analysis of the C-terminal domain of RNA polymerase II, total (Rpb1) and phosphorylated (p-Rpb1(S2)), Cyclin T1 and CDK9 protein expression in wild-type heart and liver isolated from 15-day-old (15d) and 60-day-old (60d) mice. Replicate samples are derived from independent mice. b Quantitative RT-PCR analysis of Cad, Bzw2, Pinx1, Polr3d, St6 and Cdc25a expression in wild-type (R26+/+) and R26CMER/+ heart isolated from 15-day-old (15d, Bzw2 and Cad n = 6, Cdc25a, Pinx1, Polr3d and St6n = 7) mice vs heart (Bzw2, R26+/+Pinx1 and R26CMER/+Cadn = 9, Cdc25a, Polr2d, St6, R26+/+Cad and R26CMER/+Pinx1n = 8, R26CMER/+St6n = 7, R26CMER/+Cdc25a, Polr3dn = 6) and liver (R26+/+Bzw2, Pinx1, Cad, Cdc25a and St6n = 9, R26+/+Polr2d n = 8, R26CMER/+Cadn = 10, R26CMER/+Bzw2n = 8, R26CMER/+St6n = 7, R26CMER/+Cdc25a, Pinx1, Polr3dn = 6) isolated from 60-day-old (60d) mice 4 h post administration of 4-OHT. Expression is relative to the respective wild type (R26+/+). Mean and s.d shown. One-way ANOVA with Tukey’s multiple comparisons test; 15 day R26+/+ 4-OHT vs R26CMER/+ 4-OHT : ***P = 0.001 (Cad, Bzw2, Pinx1, St6 and Cdc25a), 15 day R26+/+ 4-OHT vs R26CMER/+ 4-OHT : **P = 0.001 (Polr3d), adult heart R26+/+ 4-OHT vs R26CMER/+ 4-OHT : ***P = 0.001 (Cad), adult liver R26+/+ 4-OHT vs R26CMER/+ 4-OHT : ***P = 0.001 (Cad, Bzw2, Pinx1, St6 and Cdc25a). Replicate samples are derived from independent mice. c Heatmap showing the union of DEGs called in each tissue; adult liver (n = 3), adult heart (n = 5) and 15-day-old (15d) heart. mRNA expression fold changes (Log2) upon MycER activation are shown relative to wild type, as determined by RNA sequencing of R26CMER/+ (n = 3) tissues relative to wild type (R26+/+, n = 3) at 4 h post administration of 4-OHT. d Quantification of p-H3-positive nuclei percentage in heart (positive cardiomyocyte nuclei only) and liver (positive hepatocyte nuclei only) isolated from 15-day-old (15d) and 60-day-old (60d) wild-type (R26+/+, n = 10 15d, n = 8 60d) and R26CMER/+ (n = 4) mice 24 h post administration of tamoxifen. Means are taken from five images per mouse; mean and s.d. shown. One-way ANOVA with Tukey’s multiple comparisons test; R26+/+ 4-OHT vs R26CMER/+ 4-OHT : ***P = 0.001 (15-day heart and adult liver). Source data are provided as a Source Data file.

false

• WB

CiteAb

Western blot - Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide free (AB238940)

Western Blotting using Anti-Cyclin T1 antibody [EPR17982] - BSA and Azide freeAnti-Cyclin T1 antibody [EPR17982] - BSA and Azide free, ab238940. Publication image from Kress, T. R. et al., 2020, Nat Commun, 32286286. Legend direct from paper.

Inhibition or overexpression of P-TEFb modulates efficiency of Myc-driven transcription.a Immunoblot analysis of the C-terminal domain (CTD) of RNA Polymerase II—total (Rpb1) and phosphorylated (p-Rpb1(S5) and p-Rpb1(S2)), CDK9, Cyclin T1 and Larp7 expression in the heart, liver, lung and kidney isolated from wild-type (R26+/+) mice. The composite figure is generated from the same samples loaded across multiple blots and a representative image for GAPDH is shown. b Immunoblot analysis of the CTD of RNA Polymerase II, phosphorylated (p-Rpb1(S2)), and MycERT2 protein expression in wild type (R26+/+) or R26CMER/+ livers, isolated 4 h post administration of 4-OHT either alone (4-OHT) or in combination with 60 mg/kg AZ5576 (4-OHT + AZ5576). c Quantitative RT-PCR analysis of Smpdl3b, Cad, Gnl3 and Polr3g in wild type (R26+/+) and R26CMER/+ livers, isolated 4 h post administration of 4-OHT (n = 6 R26+/+, n = 3 R26CMER/+) either alone or in combination with 60 mg/kg AZ5576 (4-OHT + AZ5576, n = 4). Expression is relative to the respective wild type (R26+/+). Mean and s.d shown. Two-way ANOVA with Tukey’s multiple comparisons test; R26+/+ vs 4-OHT : ***P = 0.001 (Smpdl3b,Cad,Gnl3 and Polr3g), R26CMER/+ 4-OHT vs 4-OHT + AZ5576 : ***P = 0.001 (Smpdl3b, Cad and Gnl3), **P ≤ 0.01 (Polr3g). d Immunoblot analysis of Cyclin T1 and phosphorylated CTD of RNA polymerase II (p-Rpb1(S2)) in R26CMER/+ primary cardiomyocytes infected with an adenovirus encoding either GFP (Ad-GFP) or Ccnt1 (Ad-Ccnt1). Replicate samples are derived from independent primary cardiomyocyte isolations. e Quantitative RT-PCR analysis of Cad, Bzw2, Pinx1, Polr3d, St6 and Cdc25a in wild type (R26+/+, n = 5 except Pinx1 where n = 4 for Ad-GFP control) and R26CMER/+ (n = 5 except St6 where n = 4 for Ad-GFP control) primary cardiomyocytes infected with an adenovirus encoding either GFP (Ad-GFP) or Ccnt1 (Ad-Ccnt1), 4 h post addition of 100 nM 4-OHT. Expression is relative to an individual wild type (R26+/+) Ad-GFP control. Mean and s.d shown. One-way ANOVA with Tukey’s multiple comparisons test; Ad-GFP R26+/+ vs R26CMER/+ : *P = 0.05 (Cad), Ad-Ccnt1R26+/+ vs R26CMER/+ : *P = 0.05 (Cad, Pinx1, Polr3d and St6) **P = 0.01 (Bzw2 and Cdc25a). Replicate samples are derived from independent primary cardiomyocyte isolations and independent mice. Source data are provided as a Source Data file.

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