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Senescence Marker Antibody Sampler Kit

Usd: 720

Specification

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Catalog# HAK21082

Senescence Marker Antibody Sampler Kit

Application

  • WB

Reactivity

  • Human

Overview

Kit Components

Product Includes Specification Application Reactivity Mw
p16INK4a[HA601131] 20µl IHC-P,WB,IF-Cell Human Predicted band size: 16 kDa
p21[HA722685] 20µl WB,IF-Cell,IHC-P,IP Human Predicted band size: 18 kDa
Phospho-Histone H2A.X (S139)[ET1602-2] 20µl WB,IHC-P,IF-Cell,IF-Tissue Human,Mouse,Rat,Cynomolgus monkey Predicted band size: 15 kDa
Lamin B1[ET1606-27] 20µl WB,IF-Tissue,IHC-P,CUT&Tag-seq Human,Mouse,Rat Predicted band size: 66 kDa
HMGB1[ET1601-2] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC Human,Mouse,Rat Predicted band size: 25 kDa
IL-6[R1412-2] 20µl WB,IF-Cell,IHC-P,FC Human Predicted band size: 24 kDa
TNF alpha[HA722022] 20µl WB Human,Mouse Predicted band size: 26 kDa
MMP-3[ET1705-98] 20µl WB,IHC-P Human,Mouse,Rat Predicted band size: 54 kDa
Goat Anti-Rabbit IgG (H+L)[HA1001] 100µl WB,ELISA,IHC-P Rabbit
Goat Anti-Mouse IgG (H+L)[HA1006] 100µl WB,ELISA,IHC-P Mouse

Product Description

The Senescence Marker Antibody Sampler Kit provides an economical means of detecting multiple markers of cellular senescence. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Product Features

Storage Buffer

1*TBS (pH7.4), 0.05% 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

Senescence is characterized by stable stress-induced proliferative arrest and resistance to mitogenic stimuli, as well as the secretion of proteins such as cytokines, growth factors and proteases. These secreted proteins comprise the senescence-associated secretory phenotype (SASP). Senescent cells are thought to accumulate as an organism ages, and contribute to age-related diseases, including cancer, through promotion of inflammation and disruption of normal cellular function .<BR>Because there is no single biomarker that can be used to definitively identify senescent cells, researchers must rely on a collection of biomarkers commonly associated with senescence. The Senescence Marker Antibody Sampler Kit provides a collection of antibodies to commonly used biomarkers of senescence-associated cell cycle arrest (p16 INK4A, p21 Waf1/Cip1), senescence-associated DNA damage (gamma-Histone H2A.X), and the SASP (HMGB1, IL-6, TNF-alpha, MMP3). The kit also includes an antibody to Lamin B1, which is frequently reduced in senescent cells.

Data Links

Background References

1. Sun Y, Coppé JP, Lam EW. Cellular Senescence: The Sought or the Unwanted? Trends Mol Med. 2018 Oct;24(10):871-885.

2. Zhang L, Pitcher LE, Yousefzadeh MJ, Niedernhofer LJ, Robbins PD, Zhu Y. Cellular senescence: a key therapeutic target in aging and diseases. J Clin Invest. 2022 Aug 1;132(15):e158450.

Images

  • Western blot analysis of Phospho-Histone H2A.X (S139) on different lysates with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (<a href="/products/ET1602-2" style="font-weight: bold;text-decoration: underline;">ET1602-2</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa whole cell lysate<br />Lane 2: HeLa treated with 20μM Etoposide for 2 hours whole cell lysate<br />Lane 3: HeLa whole cell lysate<br />Lane 4: HeLa treated with UV for 2 hours whole cell lysate<br />Lane 5: NIH/3T3 whole cell lysate<br />Lane 6: NIH/3T3 treated with 25μM Etoposide for 5 hours whole cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 15 kDa<br />Observed band size: 15/20 kDa<br /><br />Exposure time: 53 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/ET1602-2" style="font-weight: bold;text-decoration: underline;">ET1602-2</a>) at 1/2,000 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 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Phospho-Histone H2A.X (S139) on different lysates with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/2,000 dilution.

    Lane 1: HeLa whole cell lysate
    Lane 2: HeLa treated with 20μM Etoposide for 2 hours whole cell lysate
    Lane 3: HeLa whole cell lysate
    Lane 4: HeLa treated with UV for 2 hours whole cell lysate
    Lane 5: NIH/3T3 whole cell lysate
    Lane 6: NIH/3T3 treated with 25μM Etoposide for 5 hours whole cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 15 kDa
    Observed band size: 15/20 kDa

    Exposure time: 53 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 (ET1602-2) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of HMGB1 on different lysates with Rabbit anti-HMGB1 antibody (<a href="/products/ET1601-2" style="font-weight: bold;text-decoration: underline;">ET1601-2</a>) at 1/50,000 dilution and competitor's antibody at 1/10,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: HeLa cell lysate<br />Lane 3: HCT 116 cell lysate<br />Lane 4: A549 cell lysate<br />Lane 5: Jurkat cell lysate<br />Lane 6: C2C12 cell lysate<br />Lane 7: C6 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 25 kDa<br />Observed band size: 25 kDa<br /><br />Exposure time: 21 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/ET1601-2" style="font-weight: bold;text-decoration: underline;">ET1601-2</a>) at 1/50,000 dilution and competitor's antibody at 1/10,000 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 HMGB1 on different lysates with Rabbit anti-HMGB1 antibody (ET1601-2) at 1/50,000 dilution and competitor's antibody at 1/10,000 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: HeLa cell lysate
    Lane 3: HCT 116 cell lysate
    Lane 4: A549 cell lysate
    Lane 5: Jurkat cell lysate
    Lane 6: C2C12 cell lysate
    Lane 7: C6 cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 25 kDa
    Observed band size: 25 kDa

    Exposure time: 21 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 (ET1601-2) at 1/50,000 dilution and competitor's antibody at 1/10,000 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.

  • Immunocytochemistry analysis of HeLa cells labeling HMGB1 with Rabbit anti-HMGB1 antibody (<a href="/products/ET1601-2" style="font-weight: bold;text-decoration: underline;">ET1601-2</a>) at 1/500 dilution and competitor's antibody at 1/250 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-HMGB1 antibody (<a href="/products/ET1601-2" style="font-weight: bold;text-decoration: underline;">ET1601-2</a>) at 1/500 dilution and competitor's antibody at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling HMGB1 with Rabbit anti-HMGB1 antibody (ET1601-2) at 1/500 dilution and competitor's antibody at 1/250 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-HMGB1 antibody (ET1601-2) at 1/500 dilution and competitor's antibody at 1/250 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 (M1305-2, 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.

  • Western blot analysis of HMGB1 on different lysates with Rabbit anti-HMGB1 antibody (<a href="/products/ET1601-2" style="font-weight: bold;text-decoration: underline;">ET1601-2</a>) at 1/20,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HCT 116 cell lysate (20 µg/Lane)<br />Lane 3: A549 cell lysate (20 µg/Lane)<br />Lane 4: HepG2 cell lysate (20 µg/Lane)<br />Lane 5: Jurkat cell lysate (20 µg/Lane)<br />Lane 6: Mouse thymus tissue lysate (20 µg/Lane)<br />Lane 7: Rat spleen tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 25 kDa<br />Observed band size: 25 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/ET1601-2" style="font-weight: bold;text-decoration: underline;">ET1601-2</a>) at 1/20,000 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 HMGB1 on different lysates with Rabbit anti-HMGB1 antibody (ET1601-2) at 1/20,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HCT 116 cell lysate (20 µg/Lane)
    Lane 3: A549 cell lysate (20 µg/Lane)
    Lane 4: HepG2 cell lysate (20 µg/Lane)
    Lane 5: Jurkat cell lysate (20 µg/Lane)
    Lane 6: Mouse thymus tissue lysate (20 µg/Lane)
    Lane 7: Rat spleen tissue lysate (30 µg/Lane)

    Predicted band size: 25 kDa
    Observed band size: 25 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 (ET1601-2) at 1/20,000 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 TNF alpha on different lysates with Rabbit anti-TNF alpha antibody (<a href="/products/HA722022" style="font-weight: bold;text-decoration: underline;">HA722022</a>) at 1/1,000 dilution.<br /><br />Lane 1: THP-1 cell lysate<br />Lane 2: THP-1 treated with 500ng/mL LPS for 24 hours then add 300ng/mL BFA for 20 hours cell lysate<br />Lane 3: RAW264.7 cell lysate<br />Lane 4: RAW264.7 treated with 100ng/mL LPS for 7 hours, add 300ng/mL BFA for 3 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 26 kDa<br />Observed band size: 26/50/17 kDa<br /><br />Exposure time: 1 minute;<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/HA722022" style="font-weight: bold;text-decoration: underline;">HA722022</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 TNF alpha on different lysates with Rabbit anti-TNF alpha antibody (HA722022) at 1/1,000 dilution.

    Lane 1: THP-1 cell lysate
    Lane 2: THP-1 treated with 500ng/mL LPS for 24 hours then add 300ng/mL BFA for 20 hours cell lysate
    Lane 3: RAW264.7 cell lysate
    Lane 4: RAW264.7 treated with 100ng/mL LPS for 7 hours, add 300ng/mL BFA for 3 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 26 kDa
    Observed band size: 26/50/17 kDa

    Exposure time: 1 minute;

    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 (HA722022) 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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