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Cell Cycle Regulation Antibody Sampler Kit

Usd: 720

Specification

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Catalog# HAK21039

Cell Cycle Regulation Antibody Sampler Kit

Application

  • WB

Reactivity

  • Human

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

  • Western blot analysis of Cdk2 on different lysates with Rabbit anti-Cdk2 antibody (<a href="/products/ET1602-6" style="font-weight: bold;text-decoration: underline;">ET1602-6</a>) at 1/500 dilution.<br /><br />Lane 1: A549 cell lysate, 10 µg/Lane<br />Lane 2: F9 cell lysate, 10 µg/Lane<br /><br />Predicted band size: 34 kDa<br />Observed band size: 34 kDa<br /><br />Exposure time: 2 minutes;<br /><br />12% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1602-6" style="font-weight: bold;text-decoration: underline;">ET1602-6</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    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 (<a href="/products/ET1608-61" style="font-weight: bold;text-decoration: underline;">ET1608-61</a>) at 1/2,000 dilution.<br /><br />Lane 1: MCF7 cell lysate<br />Lane 2: Jurkat cell lysate<br />Lane 3: HeLa cell lysate<br />Lane 4: Neuro-2a cell lysate<br />Lane 5: C6 cell lysate<br />Lane 6: PC-12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 22 kDa<br />Observed band size: 27 kDa<br /><br />Exposure time: 3 minutes 10 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1608-61" style="font-weight: bold;text-decoration: underline;">ET1608-61</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 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 (<a href="/products/ET1601-31" style="font-weight: bold;text-decoration: underline;">ET1601-31</a>) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution.<br /><br />Lane 1: MCF7 cell lysate, 20 µg/Lane<br />Lane 2: K-562 cell lysate (negative), 20 µg/Lane<br />Lane 3: A431 cell lysate, 20 µg/Lane<br />Lane 4: Neuro-2a cell lysate, 20 µg/Lane<br />Lane 5: NIH/3T3 cell lysate, 20 µg/Lane<br />Lane 6: C6 cell lysate, 20 µg/Lane<br />Lane 7: SH-SY5Y cell lysate, 20 µg/Lane<br /><br />Predicted band size: 34 kDa<br />Observed band size: 35 kDa<br /><br />Exposure time: 20 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1601-31" style="font-weight: bold;text-decoration: underline;">ET1601-31</a>) 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 (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) 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 (<a href="/products/ET1612-23" style="font-weight: bold;text-decoration: underline;">ET1612-23</a>) at 1/1,000 dilution.<br /><br />Lane 1: Hela cell lysate, 10 µg/Lane<br />Lane 2: MCF-7 cell lysate, 10 µg/Lane<br />Lane 3: K562 cell lysate, 10 µg/Lane<br />Lane 4: SKOV-3 cell lysate, 10 µg/Lane<br /><br />Predicted band size: 34 kDa<br />Observed band size: 34 kDa<br /><br />Exposure time: 2 minutes;<br /><br />12% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1612-23" style="font-weight: bold;text-decoration: underline;">ET1612-23</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 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 (<a href="/products/HA721960" style="font-weight: bold;text-decoration: underline;">HA721960</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: 293T cell lysate (20 µg/Lane)<br />Lane 3: MCF7 cell lysate (20 µg/Lane)<br />Lane 4: A549 cell lysate (20 µg/Lane)<br />Lane 5: Mouse skeletal muscle tissue lysate (40 µg/Lane)<br />Lane 6: Rat skeletal muscle tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 18 kDa<br />Observed band size: 18 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721960" style="font-weight: bold;text-decoration: underline;">HA721960</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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