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Loading Control Antibody Sampler Kit

Usd: 320

Specification

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Catalog# HAK21052

Loading Control Antibody Sampler Kit

Application

  • WB

Reactivity

  • Human

  • Mouse

  • Rat

Overview

Kit Components

Product Includes Specification Application Reactivity Mw
COX IV[ET1701-63] 20µl WB,IF-Cell,IF-Tissue,IHC-P,IP,FC Human,Mouse,Rat Predicted band size: 20 kDa
beta Tubulin[ET1602-4] 20µl WB,IF-Cell,IHC-P,FC Human,Mouse,Rat Predicted band size: 50 kDa
Histone H3[ET1701-64] 20µl WB,IF-Cell,IF-Tissue,IHC-P,ChIP,IP Human,Mouse,Rat Predicted band size: 15 kDa
GAPDH[HA721136] 20µl WB,IHC-P,IF-Cell,FC Human,Mouse,Rat,Escherichia coli,Zebrafish,Drosophila melanogaster Predicted band size: 36 kDa
beta Actin[HA722023] 20µl WB,IF-Cell,FC,IP Human,Mouse,Rat Predicted band size: 42 kDa
Goat Anti-Rabbit IgG (H+L)[HA1001] 100µl WB,ELISA,IHC-P Rabbit

Product Description

The Loading Control Antibody Sampler Kit contains antibodies to a variety of housekeeping proteins. The kit contains enough primary and secondary antibodies to perform two western blots per 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

Housekeeping proteins perform numerous basic functions within the cell and are constitutively expressed at high levels in a variety of tissues and cell types. Western blot analysis commonly uses housekeeping proteins such as β-actin, COX IV, GAPDH, histone H3 and the α- and β-tubulins as loading controls. <BR>Actin is a ubiquitous protein and a major component of the eukaryotic cytoskeleton. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the phosphorylation of glyceraldehyde-3-phosphate during glycolysis. Globular tubulin subunits made up of α- and β-tubulin heterodimers are the building blocks of microtubules, one of three types of cytosolic fibers that comprise the cytoskeleton. Histone proteins, including histone H3, make up the primary building block of chromatin known as nucleosomes. Modulation of the chromatin structure plays an important role in the regulation of transcription in eukaryotes. Cytochrome c oxidase (COX) is a hetero-oligomeric enzyme consisting of 13 subunits localized to the inner mitochondrial membrane .

Data Links

Background References

1. Bae BI, Hara MR, Cascio MB, Wellington CL, Hayden MR, Ross CA, Ha HC, Li XJ, Snyder SH, Sawa A. Mutant huntingtin: nuclear translocation and cytotoxicity mediated by GAPDH. Proc Natl Acad Sci U S A. 2006 Feb 28;103(9):3405-9.

2. Westermann S, Weber K. Post-translational modifications regulate microtubule function. Nat Rev Mol Cell Biol. 2003 Dec;4(12):938-47.

Images

  • Western blot analysis of beta Tubulin on different lysates with Rabbit anti-beta Tubulin antibody (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>) at 1/10,000 dilution.<br /><br />Lane 1: HeLa cell lysate (15 µg/Lane)<br />Lane 2: 293T cell lysate (15 µg/Lane)<br />Lane 3: MCF7 cell lysate (15 µg/Lane)<br />Lane 4: SH-SY5Y cell lysate (15 µg/Lane)<br />Lane 5: U-2 OS cell lysate (15 µg/Lane)<br />Lane 6: Jurkat cell lysate (15 µg/Lane)<br />Lane 7: Neuro-2a cell lysate (15 µg/Lane)<br />Lane 8: NIH/3T3 cell lysate (15 µg/Lane)<br />Lane 9: PC-12 cell lysate (15 µg/Lane)<br />Lane 10: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 11: Rat brain tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 50 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: 1 minute 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/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>) at 1/10,000 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 beta Tubulin on different lysates with Rabbit anti-beta Tubulin antibody (ET1602-4) at 1/10,000 dilution.

    Lane 1: HeLa cell lysate (15 µg/Lane)
    Lane 2: 293T cell lysate (15 µg/Lane)
    Lane 3: MCF7 cell lysate (15 µg/Lane)
    Lane 4: SH-SY5Y cell lysate (15 µg/Lane)
    Lane 5: U-2 OS cell lysate (15 µg/Lane)
    Lane 6: Jurkat cell lysate (15 µg/Lane)
    Lane 7: Neuro-2a cell lysate (15 µg/Lane)
    Lane 8: NIH/3T3 cell lysate (15 µg/Lane)
    Lane 9: PC-12 cell lysate (15 µg/Lane)
    Lane 10: Mouse brain tissue lysate (20 µg/Lane)
    Lane 11: Rat brain tissue lysate (20 µg/Lane)

    Predicted band size: 50 kDa
    Observed band size: 50 kDa

    Exposure time: 1 minute 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 (ET1602-4) at 1/10,000 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 Histone H3 on different lysates with Rabbit anti-Histone H3 antibody (<a href="/products/ET1701-64" style="font-weight: bold;text-decoration: underline;">ET1701-64</a>) at 1/20,000 dilution and competitor's antibody at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: A549 cell lysate<br />Lane 3: HT-29 cell lysate<br />Lane 4: HEK-293 cell lysate<br />Lane 5: C2C12 cell lysate<br />Lane 6: L-929 cell lysate<br />Lane 7: C6 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 15 kDa<br />Observed band size: 15 kDa<br /><br />Exposure time: 18 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/ET1701-64" style="font-weight: bold;text-decoration: underline;">ET1701-64</a>) at 1/20,000 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 Histone H3 on different lysates with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/20,000 dilution and competitor's antibody at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: A549 cell lysate
    Lane 3: HT-29 cell lysate
    Lane 4: HEK-293 cell lysate
    Lane 5: C2C12 cell lysate
    Lane 6: L-929 cell lysate
    Lane 7: C6 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 15 kDa
    Observed band size: 15 kDa

    Exposure time: 18 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 (ET1701-64) at 1/20,000 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 GAPDH on different lysates with Rabbit anti-GAPDH antibody (<a href="/products/HA721136" style="font-weight: bold;text-decoration: underline;">HA721136</a>) at 1/20,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br />Lane 3: A431 cell lysate (20 µg/Lane)<br />Lane 4: K-562 cell lysate (20 µg/Lane)<br />Lane 5: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 6: PC-12 cell lysate (20 µg/Lane)<br />Lane 7: Mouse heart tissue lysate (40 µg/Lane)<br />Lane 8: Rat heart tissue lysate (40 µg/Lane)<br />Lane 9: Human liver tissue lysate (40 µg/Lane)<br />Lane 10: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 11: Rat liver tissue lysate (40 µg/Lane)<br />Lane 12: Human kidney tissue lysate (40 µg/Lane)<br />Lane 13: Mouse kidney tissue lysate (40 µg/Lane)<br />Lane 14: Rat kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 36 kDa<br />Observed band size: 36 kDa<br /><br />Exposure time: 1 minute 2 seconds;<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/HA721136" style="font-weight: bold;text-decoration: underline;">HA721136</a>) at 1/20,000 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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of GAPDH on different lysates with Rabbit anti-GAPDH antibody (HA721136) at 1/20,000 dilution.

    Lane 1: HEK-293 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)
    Lane 3: A431 cell lysate (20 µg/Lane)
    Lane 4: K-562 cell lysate (20 µg/Lane)
    Lane 5: RAW264.7 cell lysate (20 µg/Lane)
    Lane 6: PC-12 cell lysate (20 µg/Lane)
    Lane 7: Mouse heart tissue lysate (40 µg/Lane)
    Lane 8: Rat heart tissue lysate (40 µg/Lane)
    Lane 9: Human liver tissue lysate (40 µg/Lane)
    Lane 10: Mouse liver tissue lysate (40 µg/Lane)
    Lane 11: Rat liver tissue lysate (40 µg/Lane)
    Lane 12: Human kidney tissue lysate (40 µg/Lane)
    Lane 13: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 14: Rat kidney tissue lysate (40 µg/Lane)

    Predicted band size: 36 kDa
    Observed band size: 36 kDa

    Exposure time: 1 minute 2 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 (HA721136) at 1/20,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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 beta Actin on different lysates with Rabbit anti-beta Actin antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>) at 1/20,000 dilution.<br /><br />Lane 1: HeLa cell lysate (10 µg/Lane)<br />Lane 2: HepG2 cell lysate (10 µg/Lane)<br />Lane 3: MCF7 cell lysate (10 µg/Lane)<br />Lane 4: A431 cell lysate (10 µg/Lane)<br />Lane 5: Jurkat cell lysate (10 µg/Lane)<br />Lane 6: HEK-293 cell lysate (10 µg/Lane)<br />Lane 7: RAW264.7 cell lysate (10 µg/Lane)<br />Lane 8: C2C12 cell lysate (10 µg/Lane)<br />Lane 9: PC-12 cell lysate (10 µg/Lane)<br />Lane 10: Mouse testis tissue lysate (10 µg/Lane)<br />Lane 11: Mouse spleen tissue lysate (10 µg/Lane)<br />Lane 12: Rat testis tissue lysate (10 µg/Lane)<br />Lane 13: Rat kidney tissue lysate (10 µg/Lane)<br /><br />Predicted band size: 42 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 3 seconds;<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/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</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 beta Actin on different lysates with Rabbit anti-beta Actin antibody (HA722023) at 1/20,000 dilution.

    Lane 1: HeLa cell lysate (10 µg/Lane)
    Lane 2: HepG2 cell lysate (10 µg/Lane)
    Lane 3: MCF7 cell lysate (10 µg/Lane)
    Lane 4: A431 cell lysate (10 µg/Lane)
    Lane 5: Jurkat cell lysate (10 µg/Lane)
    Lane 6: HEK-293 cell lysate (10 µg/Lane)
    Lane 7: RAW264.7 cell lysate (10 µg/Lane)
    Lane 8: C2C12 cell lysate (10 µg/Lane)
    Lane 9: PC-12 cell lysate (10 µg/Lane)
    Lane 10: Mouse testis tissue lysate (10 µg/Lane)
    Lane 11: Mouse spleen tissue lysate (10 µg/Lane)
    Lane 12: Rat testis tissue lysate (10 µg/Lane)
    Lane 13: Rat kidney tissue lysate (10 µg/Lane)

    Predicted band size: 42 kDa
    Observed band size: 42 kDa

    Exposure time: 3 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 (HA722023) 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 COX IV on different lysates with Rabbit anti-COX IV antibody (<a href="/products/ET1701-63" style="font-weight: bold;text-decoration: underline;">ET1701-63</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HepG2 cell lysate (20 µg/Lane)<br />Lane 3: Mouse heart tissue lysate (30 µg/Lane)<br />Lane 4: Rat heart tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 20 kDa<br />Observed band size: 15 kDa<br /><br />Exposure time: 10 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/ET1701-63" style="font-weight: bold;text-decoration: underline;">ET1701-63</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 COX IV on different lysates with Rabbit anti-COX IV antibody (ET1701-63) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HepG2 cell lysate (20 µg/Lane)
    Lane 3: Mouse heart tissue lysate (30 µg/Lane)
    Lane 4: Rat heart tissue lysate (30 µg/Lane)

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

    Exposure time: 10 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 (ET1701-63) 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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