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Mitochondrial Marker Antibody Sampler Kit

Usd: 720

Specification

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Catalog# HAK21041

Mitochondrial Marker Antibody Sampler Kit

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

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
Cytochrome C[ET1610-60] 20µl WB,IF-Tissue,IHC-P,IP,IF-Cell Human,Mouse,Rat Predicted band size: 12 kDa
HSP60[ET1609-45] 20µl WB,IHC-P,IF-Cell,IP Human,Mouse,Rat Predicted band size: 61 kDa
Prohibitin[ET1602-31] 20µl WB,IF-Cell,IHC-P,IP,FC Human,Mouse,Rat Predicted band size: 30 kDa
PDHA1[ET1702-75] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC(Intra) Human,Mouse,Rat Predicted band size: 43 kDa
SDHA[ET1703-40] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC Human,Mouse,Rat,Zebrafish Predicted band size: 73 kDa
Superoxide Dismutase 1[ET1702-36] 20µl WB,IF-Cell,IHC-P,FC Human,Mouse,Rat Predicted band size: 16 kDa
VDAC1[ET1601-20] 20µl WB,IF-Cell,IHC-P,FC,IF-Tissue Human,Mouse,Rat Predicted band size: 31 kDa
Goat Anti-Rabbit IgG (H+L)[HA1001] 100µl WB,ELISA,IHC-P Rabbit

Product Description

The Mitochondrial Marker Antibody Sampler Kit provides an economical means to evaluate relevant mitochondial proteins. This kit contains enough primary antibody to perform two western blots per primary.

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

Cytochrome c oxidase (COX) is a hetero-oligomeric enzyme consisting of 13 subunits localized to the inner mitochondrial membrane. Cytochrome c is a well conserved electron-transport protein and is part of the respiratory chain localized to the mitochondrial intermembrane space. <br>HSP60 has primarily been known as a mitochondrial protein that is important for folding key proteins after import into the mitochondria. In the mitochondria, prohibitins (PHB1) mainly exist as membrane-bound ring complexes and function as chaperones maintaining mitochondrial protein stability during protein synthesis and transportation. In mammalian cells, the pyruvate dehydrogenase complex is located in the mitochondrial matrix. Succinate dehydrogenase (SDH), also known as Complex II or succinate quinone oxidoreductase, is a key component of the citric acid cycle and the electron transport chain. SOD1 is ubiquitously expressed and is localized in the cytosol, nucleus, and mitochondrial intermembrane space. <br>Voltage-dependent anion channel (VDAC), ubiquitously expressed and located in the outer mitochondrial membrane, is generally thought to be the primary means by which metabolites diffuse in and out of the mitochondria.

Data Links

Background References

1. Schägger H. Respiratory chain supercomplexes of mitochondria and bacteria. Biochim Biophys Acta. 2002 Sep 10;1555(1-3):154-9.

2. Tatsuta T, Model K, Langer T. Formation of membrane-bound ring complexes by prohibitins in mitochondria. Mol Biol Cell. 2005 Jan;16(1):248-59.

3. Craigen WJ, Graham BH. Genetic strategies for dissecting mammalian and Drosophila voltage-dependent anion channel functions. J Bioenerg Biomembr. 2008 Jun;40(3):207-12.

Images

  • 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.

  • Western blot analysis of Cytochrome C on different lysates with Rabbit anti-Cytochrome C antibody (<a href="/products/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (15 µg/Lane)<br />Lane 2: Jurkat cell lysate (15 µg/Lane)<br />Lane 3: C6 cell lysate (15 µg/Lane)<br />Lane 4: Human kidney tissue lysate (20 µg/Lane)<br />Lane 5: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 6: Rat brain tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 12 kDa<br />Observed band size: 14 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/ET1610-60" style="font-weight: bold;text-decoration: underline;">ET1610-60</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 Cytochrome C on different lysates with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (15 µg/Lane)
    Lane 2: Jurkat cell lysate (15 µg/Lane)
    Lane 3: C6 cell lysate (15 µg/Lane)
    Lane 4: Human kidney tissue lysate (20 µg/Lane)
    Lane 5: Mouse brain tissue lysate (20 µg/Lane)
    Lane 6: Rat brain tissue lysate (20 µg/Lane)

    Predicted band size: 12 kDa
    Observed band size: 14 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 (ET1610-60) 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 Hsp60 on different lysates with Rabbit anti-Hsp60 antibody (<a href="/products/ET1609-45" style="font-weight: bold;text-decoration: underline;">ET1609-45</a>) at 1/100,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HepG2 cell lysate<br />Lane 3: SW480 cell lysate<br />Lane 4: Jurkat cell lysate<br />Lane 5: PANC-1 cell lysate<br />Lane 6: F9 cell lysate<br />Lane 7: NIH/3T3 cell lysate<br />Lane 8: PC-12 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 61 kDa<br />Observed band size: 61 kDa<br /><br />Exposure time: 46 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/ET1609-45" style="font-weight: bold;text-decoration: underline;">ET1609-45</a>) at 1/100,000 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 Hsp60 on different lysates with Rabbit anti-Hsp60 antibody (ET1609-45) at 1/100,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HepG2 cell lysate
    Lane 3: SW480 cell lysate
    Lane 4: Jurkat cell lysate
    Lane 5: PANC-1 cell lysate
    Lane 6: F9 cell lysate
    Lane 7: NIH/3T3 cell lysate
    Lane 8: PC-12 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 61 kDa
    Observed band size: 61 kDa

    Exposure time: 46 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 (ET1609-45) at 1/100,000 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 Prohibitin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>, 1/500) was used in 5% BSA 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 1:5,000 dilution was used for 1 hour at room temperature.<br /><span style="font-weight: bold;">Positive control:</span><br />Lane 1: Jurkat cell lysate<br />Lane 2: Mouse kidney tissue lysate<br />Lane 3: 293 cell lysate<br />Lane 4: HepG2 cell lysate

    Western blot analysis of Prohibitin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1602-31, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
    Positive control:
    Lane 1: Jurkat cell lysate
    Lane 2: Mouse kidney tissue lysate
    Lane 3: 293 cell lysate
    Lane 4: HepG2 cell lysate

  • Western blot analysis of Superoxide Dismutase 1 on different lysates with Rabbit anti-Superoxide Dismutase 1 antibody (<a href="/products/ET1702-36" style="font-weight: bold;text-decoration: underline;">ET1702-36</a>) at 1/1,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 2: C2C12 cell lysate (20 µg/Lane)<br />Lane 3: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 4: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 5: Mouse hippocampus tissue lysate (40 µg/Lane)<br />Lane 6: Mouse stomach tissue lysate (40 µg/Lane)<br />Lane 7: Mouse smooth muscle tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 16 kDa<br />Observed band size: 16 kDa<br /><br />Exposure time: 24 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/ET1702-36" style="font-weight: bold;text-decoration: underline;">ET1702-36</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 Superoxide Dismutase 1 on different lysates with Rabbit anti-Superoxide Dismutase 1 antibody (ET1702-36) at 1/1,000 dilution.

    Lane 1: RAW264.7 cell lysate (20 µg/Lane)
    Lane 2: C2C12 cell lysate (20 µg/Lane)
    Lane 3: Mouse brain tissue lysate (40 µg/Lane)
    Lane 4: Mouse liver tissue lysate (40 µg/Lane)
    Lane 5: Mouse hippocampus tissue lysate (40 µg/Lane)
    Lane 6: Mouse stomach tissue lysate (40 µg/Lane)
    Lane 7: Mouse smooth muscle tissue lysate (40 µg/Lane)

    Predicted band size: 16 kDa
    Observed band size: 16 kDa

    Exposure time: 24 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 (ET1702-36) 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.

  • Western blot analysis of VDAC1 on different lysates with Rabbit anti-VDAC1 antibody (<a href="/products/ET1601-20" style="font-weight: bold;text-decoration: underline;">ET1601-20</a>) at 1/10,000 dilution and competitor's antibody at 1/10,000 dilution.<br /><br />Lane 1: MDA-MB-231 cell lysate, 15 µg/Lane<br />Lane 2: HeLa cell lysate, 15 µg/Lane<br />Lane 3: K-562 cell lysate, 15 µg/Lane<br />Lane 4: NIH/3T3 cell lysate, 15 µg/Lane<br />Lane 5: PC-12 cell lysate, 15 µg/Lane<br />Lane 6: Mouse kidney tissue lysate, 15 µg/Lane<br /><br />Predicted band size: 31 kDa<br />Observed band size: 31 kDa<br /><br />Exposure time: 14 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/ET1601-20" style="font-weight: bold;text-decoration: underline;">ET1601-20</a>) at 1/10,000 dilution and competitor's antibody at 1/10,000 dilution were 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 VDAC1 on different lysates with Rabbit anti-VDAC1 antibody (ET1601-20) at 1/10,000 dilution and competitor's antibody at 1/10,000 dilution.

    Lane 1: MDA-MB-231 cell lysate, 15 µg/Lane
    Lane 2: HeLa cell lysate, 15 µg/Lane
    Lane 3: K-562 cell lysate, 15 µg/Lane
    Lane 4: NIH/3T3 cell lysate, 15 µg/Lane
    Lane 5: PC-12 cell lysate, 15 µg/Lane
    Lane 6: Mouse kidney tissue lysate, 15 µg/Lane

    Predicted band size: 31 kDa
    Observed band size: 31 kDa

    Exposure time: 14 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 (ET1601-20) at 1/10,000 dilution and competitor's antibody at 1/10,000 dilution were 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.

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

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