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ER Stress Antibody Sampler Kit

Usd: 640

Specification

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Catalog# HAK21037

ER Stress Antibody Sampler Kit

Application

  • WB

Reactivity

  • Human

Overview

Kit Components

Product Includes Specification Application Reactivity Mw
GRP78 / BIP[HA601076] 20µl WB,IHC-P,IF-Cell Human,Mouse,Rat Predicted band size: 72 kDa
Calnexin[ET1611-86] 20µl WB,IP,IHC-P,IF-Tissue Human,Mouse,Rat Predicted band size: 68 kDa
ERO1L[HA722818] 20µl WB,IHC-P Human,Mouse,Rat Predicted band size: 54 kDa
IRE1[HA723225] 20µl WB Human,Mouse,Rat,Monkey Predicted band size: 110 kDa
P4HB[ET7110-92] 20µl WB,IF-Cell,IHC-P,FC Human,Mouse,Rat,Monkey Predicted band size: 57 kDa
DDIT3[ET1703-05] 20µl WB,FC Human,Mouse,Rat Predicted band size: 19 kDa
PERK[HA721510] 20µl WB Human Predicted band size: 125 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 ER Stress Sampler Kit contains reagents to investigate ER stress within the cell. The kit contains enough primary and secondary antibodies to perform two Western blot experiments per 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

Secretory and transmembrane proteins are synthesized on polysomes and translocate into the endoplasmic reticulum (ER).The ER contains a pool of molecular chaperone proteins including calnexin, BiP and protein disulfide isomerase (PDI).</br>Calnexin is an ER membrane, calcium-binding protein that retains newly synthesized glycoproteins inside the ER to ensure proper folding and quality control.Irregular protein folding within the ER increases BiP synthesis, which binds misfolded proteins to prevent them from forming aggregates and to assist them to refold properly.</br>PDI catalyzes the formation and isomerization of disulfide bonds required for a protein to reach its native state.Ero1-Lα is an ER membrane-associated N-glycoprotein that promotes oxidative protein folding.This is regulated by proteins such asthe membrane-bound transcription factor protease site 2 (MBTPS2) and the serine/threonine kinase IRE1.The PERK eIF2α kinase is an ER resident transmembrane protein that couples ER stress signals to translation inhibition.During ER stress, the level of CHOP expression is elevated and CHOP functions to mediate programmed cell death.

Data Links

Background References

1. Williams DB. Beyond lectins: the calnexin/calreticulin chaperone system of the endoplasmic reticulum. J Cell Sci. 2006 Feb 15;119(Pt 4):615-23.

2. Di Conza G, Ho PC. ER Stress Responses: An Emerging Modulator for Innate Immunity. Cells. 2020 Mar 12;9(3):695.

3. Rellmann Y, Eidhof E, Dreier R. Review: ER stress-induced cell death in osteoarthritic cartilage. Cell Signal. 2021 Feb;78:109880.

4. Kaufman RJ, Scheuner D, Schröder M, Shen X, Lee K, Liu CY, Arnold SM. The unfolded protein response in nutrient sensing and differentiation. Nat Rev Mol Cell Biol. 2002 Jun;3(6):411-21.

Images

  • Western blot analysis of GRP78 / BIP on different lysates with Mouse anti-GRP78 / BIP antibody (<a href="/products/HA601076" style="font-weight: bold;text-decoration: underline;">HA601076</a>) at 1/1,000 dilution.<br /><br />Lane 1: MCF7 cell lysate (15 µg/Lane)<br />Lane 2: HepG2 cell lysate (15 µg/Lane)<br />Lane 3: Mouse brain tissue lysate (30 µg/Lane)<br />Lane 4: U-87 MG cell lysate (30 µg/Lane)<br />Lane 5: RAW264.7 cell lysate (30 µg/Lane)<br />Lane 6: RAW264.7 treated with 300nM Thapsigargin for 18 hours cell lysate (30 µg/Lane)<br />Lane 7: Mouse liver tissue lysate (30 µg/Lane)<br />Lane 8: Rat liver tissue lysate (30 µg/Lane)<br />Lane 9: Rat pancreas tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 72 kDa<br />Observed band size: 72 kDa<br /><br />Exposure time: Lane 1-3: 11 seconds; Lane 4-9: 4 seconds; ECL: K1801;<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/HA601076" style="font-weight: bold;text-decoration: underline;">HA601076</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of GRP78 / BIP on different lysates with Mouse anti-GRP78 / BIP antibody (HA601076) at 1/1,000 dilution.

    Lane 1: MCF7 cell lysate (15 µg/Lane)
    Lane 2: HepG2 cell lysate (15 µg/Lane)
    Lane 3: Mouse brain tissue lysate (30 µg/Lane)
    Lane 4: U-87 MG cell lysate (30 µg/Lane)
    Lane 5: RAW264.7 cell lysate (30 µg/Lane)
    Lane 6: RAW264.7 treated with 300nM Thapsigargin for 18 hours cell lysate (30 µg/Lane)
    Lane 7: Mouse liver tissue lysate (30 µg/Lane)
    Lane 8: Rat liver tissue lysate (30 µg/Lane)
    Lane 9: Rat pancreas tissue lysate (30 µg/Lane)

    Predicted band size: 72 kDa
    Observed band size: 72 kDa

    Exposure time: Lane 1-3: 11 seconds; Lane 4-9: 4 seconds; 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 (HA601076) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of Hela cells labeling GRP78 / BIP with Mouse anti-GRP78 / BIP antibody (<a href="/products/HA601076" style="font-weight: bold;text-decoration: underline;">HA601076</a>) at 1/200 dilution.<br /><br />Cells were fixed in 100% methanol for 10 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-GRP78 / BIP antibody (<a href="/products/HA601076" style="font-weight: bold;text-decoration: underline;">HA601076</a>) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of Hela cells labeling GRP78 / BIP with Mouse anti-GRP78 / BIP antibody (HA601076) at 1/200 dilution.

    Cells were fixed in 100% methanol for 10 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-GRP78 / BIP antibody (HA601076) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Western blot analysis of Calnexin on different lysates with Rabbit anti-Calnexin antibody (<a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HepG2 cell lysate<br />Lane 3: MCF7 cell lysate<br />Lane 4: PANC-1 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 68 kDa<br />Observed band size: 100 kDa<br /><br />Exposure time: 2 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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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 Calnexin on different lysates with Rabbit anti-Calnexin antibody (ET1611-86) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HepG2 cell lysate
    Lane 3: MCF7 cell lysate
    Lane 4: PANC-1 cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 68 kDa
    Observed band size: 100 kDa

    Exposure time: 2 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 (ET1611-86) 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 DDIT3 on different lysates with Rabbit anti-DDIT3 antibody (<a href="/products/ET1703-05" style="font-weight: bold;text-decoration: underline;">ET1703-05</a>) at 1/5,000 dilution.<br /><br />Lane 1: SW480 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br /><br />Predicted band size: 19 kDa<br />Observed band size: 27 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/ET1703-05" style="font-weight: bold;text-decoration: underline;">ET1703-05</a>) at 1/5,000 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 DDIT3 on different lysates with Rabbit anti-DDIT3 antibody (ET1703-05) at 1/5,000 dilution.

    Lane 1: SW480 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)

    Predicted band size: 19 kDa
    Observed band size: 27 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 (ET1703-05) at 1/5,000 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 PERK on different lysates with Rabbit anti-PERK antibody (<a href="/products/HA721510" style="font-weight: bold;text-decoration: underline;">HA721510</a>) at 1/1,000 dilution.<br /><br />Lane 1: MCF7 cell lysate (20 µg/Lane)<br />Lane 2: HepG2 cell lysate (20 µg/Lane)<br />Lane 3: HEK-293 cell lysate (20 µg/Lane)<br />Lane 4: HeLa cell lysate (20 µg/Lane)<br />Lane 5: A549 cell lysate (20 µg/Lane)<br />Lane 6: U-87 MG cell lysate (20 µg/Lane)<br />Lane 7: K-562 cell lysate (16 µg/Lane)<br /><br />Predicted band size: 125 kDa<br />Observed band size: 140 kDa<br /><br />Exposure time: 2 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/HA721510" style="font-weight: bold;text-decoration: underline;">HA721510</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 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of PERK on different lysates with Rabbit anti-PERK antibody (HA721510) at 1/1,000 dilution.

    Lane 1: MCF7 cell lysate (20 µg/Lane)
    Lane 2: HepG2 cell lysate (20 µg/Lane)
    Lane 3: HEK-293 cell lysate (20 µg/Lane)
    Lane 4: HeLa cell lysate (20 µg/Lane)
    Lane 5: A549 cell lysate (20 µg/Lane)
    Lane 6: U-87 MG cell lysate (20 µg/Lane)
    Lane 7: K-562 cell lysate (16 µg/Lane)

    Predicted band size: 125 kDa
    Observed band size: 140 kDa

    Exposure time: 2 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 (HA721510) 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 1:100,000 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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