Cell Cycle Regulation Antibody Sampler Kit
Catalog# HAK21039
Cell Cycle Regulation Antibody Sampler Kit
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WB
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Human
General Overview
Kit Components
| Product Includes | Specification | Application | Reactivity | Mw |
|---|---|---|---|---|
| Cdk2[ET1602-6] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,IP,FC | Human,Mouse | Predicted band size: 34 kDa |
| p27 KIP 1[ET1608-61] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,FC,IP | Human,Mouse,Rat | Predicted band size: 22 kDa |
| Cyclin D1[ET1601-31] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,IP,FC | Human,Mouse,Rat | Predicted band size: 34 kDa |
| Cdk6[ET1612-3] | 20µl | IF-Cell,IF-Tissue,IHC-P,FC,WB | Human | Predicted band size: 37 kDa |
| Cyclin D3[ET1612-4] | 20µl | WB | Human,Mouse,Rat | Predicted band size: 33 kDa |
| p21[HA722685] | 20µl | WB,IF-Cell,IHC-P,IP | Human | Predicted band size: 18 kDa |
| Cdk4[ET1612-23] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,FC | Human,Mouse,Rat | Predicted band size: 34 kDa |
| p18 INK4c[HA721960] | 20µl | WB,IHC-P | Human,Mouse,Rat | Predicted band size: 18 kDa |
| Goat Anti-Rabbit IgG (H+L)[HA1001] | 100µl | WB,ELISA,IHC-P | Rabbit |
Product Description
Cell Cycle Regulation Antibody Sampler kit offers an economical way of detecting eight integral cell cycle regulation proteins. The kit contains enough primary and secondary antibodies to perform two western blot experiments with each primary antibody.
Product Features
Storage Buffer
1*TBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Background
Eukaryotic cell cycle progression is dependent, in part, on the tightly regulated activity of cyclin dependent kinases (CDKs). Cyclin D/CDK4/6 activity occurs in mid-late G1 phase, upstream of CDK2/cyclin E activity. <br>Cyclin D is degraded through the ubiquitin proteasome pathway in the absence of mitogenic signaling. p27/Kip1, p57 Kip2 and p21 Waf1/Cip1 are members of the Cip/Kip family of cyclin-dependent kinase inhibitors. They form heterotrimeric complexes with cyclins and CDKs, inhibiting kinase activity and blocking progression through G1/S phase . <BR>Levels of p27 are upregulated in quiescent cells and in cells treated with negative cell cycle regulators. p27 nuclear localization is controlled by Akt-dependent phosphorylation at Thr157. The inhibitors of CDK4 (INK4) family include p15 INK4B, p16 INK4A, p18 INK4C, and p19 INK4D. All INK4 proteins selectively inhibit CDK4/6 activity, either in a binary complex, or in a ternary complex including cyclin D, resulting in inhibition of cell division.
Data Links
Background References
1. Shin I, Yakes FM, Rojo F, Shin NY, Bakin AV, Baselga J, Arteaga CL. PKB/Akt mediates cell-cycle progression by phosphorylation of p27(Kip1) at threonine 157 and modulation of its cellular localization. Nat Med. 2002 Oct;8(10):1145-52.
2. Cheng M, Olivier P, Diehl JA, Fero M, Roussel MF, Roberts JM, Sherr CJ. The p21(Cip1) and p27(Kip1) CDK \'inhibitors\' are essential activators of cyclin D-dependent kinases in murine fibroblasts. EMBO J. 1999 Mar 15;18(6):1571-83.
3. Sheaff RJ, Singer JD, Swanger J, Smitherman M, Roberts JM, Clurman BE. Proteasomal turnover of p21Cip1 does not require p21Cip1 ubiquitination. Mol Cell. 2000 Feb;5(2):403-10.
4. Hirai H, Roussel MF, Kato JY, Ashmun RA, Sherr CJ. Novel INK4 proteins, p19 and p18, are specific inhibitors of the cyclin D-dependent kinases CDK4 and CDK6. Mol Cell Biol. 1995 May;15(5):2672-81.
Images
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Western blot analysis of Cdk2 on different lysates with Rabbit anti-Cdk2 antibody (ET1602-6) at 1/500 dilution.
Lane 1: A549 cell lysate, 10 µg/Lane
Lane 2: F9 cell lysate, 10 µg/Lane
Predicted band size: 34 kDa
Observed band size: 34 kDa
Exposure time: 2 minutes;
12% 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 (ET1602-6) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of p27 KIP 1 on different lysates with Rabbit anti-p27 KIP 1 antibody (ET1608-61) at 1/2,000 dilution.
Lane 1: MCF7 cell lysate
Lane 2: Jurkat cell lysate
Lane 3: HeLa cell lysate
Lane 4: Neuro-2a cell lysate
Lane 5: C6 cell lysate
Lane 6: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 22 kDa
Observed band size: 27 kDa
Exposure time: 3 minutes 10 seconds;
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 (ET1608-61) 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. -
Western blot analysis of Cyclin D1 on different lysates with Rabbit anti-Cyclin D1 antibody (ET1601-31) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution.
Lane 1: MCF7 cell lysate, 20 µg/Lane
Lane 2: K-562 cell lysate (negative), 20 µg/Lane
Lane 3: A431 cell lysate, 20 µg/Lane
Lane 4: Neuro-2a cell lysate, 20 µg/Lane
Lane 5: NIH/3T3 cell lysate, 20 µg/Lane
Lane 6: C6 cell lysate, 20 µg/Lane
Lane 7: SH-SY5Y cell lysate, 20 µg/Lane
Predicted band size: 34 kDa
Observed band size: 35 kDa
Exposure time: 20 seconds;
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 (ET1601-31) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution were 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. -
Western blot analysis of Cdk4 on different lysates with Rabbit anti-Cdk4 antibody (ET1612-23) at 1/1,000 dilution.
Lane 1: Hela cell lysate, 10 µg/Lane
Lane 2: MCF-7 cell lysate, 10 µg/Lane
Lane 3: K562 cell lysate, 10 µg/Lane
Lane 4: SKOV-3 cell lysate, 10 µg/Lane
Predicted band size: 34 kDa
Observed band size: 34 kDa
Exposure time: 2 minutes;
12% 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 (ET1612-23) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of p18 INK4c on different lysates with Rabbit anti-p18 INK4c antibody (HA721960) at 1/1,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: 293T cell lysate (20 µg/Lane)
Lane 3: MCF7 cell lysate (20 µg/Lane)
Lane 4: A549 cell lysate (20 µg/Lane)
Lane 5: Mouse skeletal muscle tissue lysate (40 µg/Lane)
Lane 6: Rat skeletal muscle tissue lysate (40 µg/Lane)
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 3 minutes;
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 (HA721960) at 1/1,000 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.
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