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Reactive Exosome Marker Antibody Sampler Kit

Usd: 560

Specification

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Catalog# HAK21058

Reactive Exosome Marker Antibody Sampler Kit

Application

  • WB

Reactivity

  • Human

  • Mouse

  • Rat

Overview

Kit Components

Product Includes Specification Application Reactivity Mw
CD9[ET1601-9] 20µl WB,IF-Cell,IHC-P,IP,FC Human,Mouse,Rat Predicted band size: 25 kDa
TAPA1/CD81[ET1611-87] 20µl WB,IF-Cell,IHC-P,FC Human,Mouse,Rat Predicted band size: 26 kDa
TSG101[ET1701-59] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC Human,Mouse,Rat Predicted band size: 44 kDa
ALIX[ET1705-74] 20µl WB,IHC-P,FC,IF-Cell Human,Mouse,Rat Predicted band size: 96 kDa
Flotillin 1[ET7107-82] 20µl WB,IHC-P Human,Mouse,Rat Predicted band size: 47 kDa
Hsp70[ET1601-11] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC Human,Mouse,Rat Predicted band size: 70 kDa
Goat Anti-Rabbit IgG (H+L)[HA1001] 100µl WB,ELISA,IHC-P Rabbit

Product Description

The Human Reactive Exosome Marker Antibody Sampler Kit provides an economical means of analyzing proteins that can be present on exosomes. The kit includes enough antibodies 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

Exosomes are small (30-150 nm) membrane-bound vesicles that are secreted by various cell types under normal and pathological conditions.There are protein markers that appear with high frequency. Tetraspanins CD9, CD81, and CD63 appear in exosomes and have been the target of immune-affinity approaches of exosome isolation. <BR>Flotillin-1 is a lipid raft-associated integral membrane protein that is incorporated into exosomes. Alix regulates cellular processes, such as endocytic membrane trafficking and cell adhesion through interactions with ESCRT proteins including endophilins, and CIN85 (Cbl-interacting protein of 85 kDa), and plays a role in exosome biogenesis. Syntenin-1 (MDA9, SDCBP) is a member of the PDZ family of proteins that functions as a scaffold adaptor protein regulating numerous signal transduction pathways.Tumor susceptibility gene 101 (TSG101) is a fundamental component of the ESCRT complex I involved in regulating the trafficking of proteins throughout the endosomal compartment. HSP70 may also be secreted through exosomes .

Data Links

Background References

1. Sidhom K, Obi PO, Saleem A. A Review of Exosomal Isolation Methods: Is Size Exclusion Chromatography the Best Option? Int J Mol Sci. 2020 Sep 4;21(18):6466.

2. Pradhan AK, Maji S, Das SK, Emdad L, Sarkar D, Fisher PB. MDA-9/Syntenin/SDCBP: new insights into a unique multifunctional scaffold protein. Cancer Metastasis Rev. 2020 Sep;39(3):769-781.

Images

  • Western blot analysis of CD9 on different lysates with Rabbit anti-CD9 antibody (<a href="/products/ET1601-9" style="font-weight: bold;text-decoration: underline;">ET1601-9</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: K-562 cell lysate (20 µg/Lane)<br />Lane 3: MCF7 cell lysate (20 µg/Lane)<br />Lane 4: HCT 116 cell lysate (20 µg/Lane)<br />Lane 5: HepG2 cell lysate (20 µg/Lane)<br />Lane 6: SK-MEL-28 cell lysate (20 µg/Lane)<br />Lane 7: A375 cell lysate (20 µg/Lane)<br />Lane 8: B16-F1 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 25 kDa<br />Observed band size: 23 kDa<br /><br />Exposure time: 3 minutes 30 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/ET1601-9" style="font-weight: bold;text-decoration: underline;">ET1601-9</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 CD9 on different lysates with Rabbit anti-CD9 antibody (ET1601-9) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: K-562 cell lysate (20 µg/Lane)
    Lane 3: MCF7 cell lysate (20 µg/Lane)
    Lane 4: HCT 116 cell lysate (20 µg/Lane)
    Lane 5: HepG2 cell lysate (20 µg/Lane)
    Lane 6: SK-MEL-28 cell lysate (20 µg/Lane)
    Lane 7: A375 cell lysate (20 µg/Lane)
    Lane 8: B16-F1 cell lysate (20 µg/Lane)

    Predicted band size: 25 kDa
    Observed band size: 23 kDa

    Exposure time: 3 minutes 30 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 (ET1601-9) 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 TSG101 on different lysates with Rabbit anti-TSG101 antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: K-562 cell lysate (20 µg/Lane)<br />Lane 3: MCF7 cell lysate (20 µg/Lane)<br />Lane 4: Jurkat cell lysate (20 µg/Lane)<br />Lane 5: C2C12 cell lysate (20 µg/Lane)<br />Lane 6: PC-12 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 44 kDa<br />Observed band size: 44/47 kDa<br /><br />Exposure time: 3 minutes 20 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/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/2,000 dilution and competitor's antibody at 1/1,000 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 TSG101 on different lysates with Rabbit anti-TSG101 antibody (ET1701-59) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: K-562 cell lysate (20 µg/Lane)
    Lane 3: MCF7 cell lysate (20 µg/Lane)
    Lane 4: Jurkat cell lysate (20 µg/Lane)
    Lane 5: C2C12 cell lysate (20 µg/Lane)
    Lane 6: PC-12 cell lysate (20 µg/Lane)

    Predicted band size: 44 kDa
    Observed band size: 44/47 kDa

    Exposure time: 3 minutes 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 (ET1701-59) at 1/2,000 dilution and competitor's antibody at 1/1,000 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 ALIX on different lysates with Rabbit anti-ALIX antibody (<a href="/products/ET1705-74" style="font-weight: bold;text-decoration: underline;">ET1705-74</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 2: PC-12 cell lysate (20 µg/Lane)<br />Lane 3: HEK-293 cell lysate (20 µg/Lane)<br />Lane 4: K-562 cell lysate (20 µg/Lane)<br />Lane 5: Mouse liver tissue lysate (30 µg/Lane)<br />Lane 6: Rat liver tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 96 kDa<br />Observed band size: 80-100 kDa<br /><br />Exposure time: 6 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/ET1705-74" style="font-weight: bold;text-decoration: underline;">ET1705-74</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 ALIX on different lysates with Rabbit anti-ALIX antibody (ET1705-74) at 1/1,000 dilution.

    Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 2: PC-12 cell lysate (20 µg/Lane)
    Lane 3: HEK-293 cell lysate (20 µg/Lane)
    Lane 4: K-562 cell lysate (20 µg/Lane)
    Lane 5: Mouse liver tissue lysate (30 µg/Lane)
    Lane 6: Rat liver tissue lysate (30 µg/Lane)

    Predicted band size: 96 kDa
    Observed band size: 80-100 kDa

    Exposure time: 6 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 (ET1705-74) 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 Flotillin 1 on different lysates with Rabbit anti-Flotillin 1 antibody (<a href="/products/ET7107-82" style="font-weight: bold;text-decoration: underline;">ET7107-82</a>) at 1/1,000 dilution.<br /><br />Lane 1: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 2: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 3: Mouse kidney tissue lysate (40 µg/Lane)<br />Lane 4: Mouse lung tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 47 kDa<br />Observed band size: 47 kDa<br /><br />Exposure time: 59 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/ET7107-82" style="font-weight: bold;text-decoration: underline;">ET7107-82</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 Flotillin 1 on different lysates with Rabbit anti-Flotillin 1 antibody (ET7107-82) at 1/1,000 dilution.

    Lane 1: Neuro-2a cell lysate (20 µg/Lane)
    Lane 2: Mouse brain tissue lysate (40 µg/Lane)
    Lane 3: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 4: Mouse lung tissue lysate (40 µg/Lane)

    Predicted band size: 47 kDa
    Observed band size: 47 kDa

    Exposure time: 59 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 (ET7107-82) 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 Hsp70 on different lysates with Rabbit anti-Hsp70 antibody (<a href="/products/ET1601-11" style="font-weight: bold;text-decoration: underline;">ET1601-11</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: A549 cell lysate<br />Lane 3: MCF7 cell lysate<br />Lane 4: HCT 116 cell lysate<br />Lane 5: Mouse brain tissue lysate<br />Lane 6: Mouse testis tissue lysate<br />Lane 7: Rat testis tissue lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 70 kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: Lane 1-4: 43 seconds; Lane 5-7: 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/ET1601-11" style="font-weight: bold;text-decoration: underline;">ET1601-11</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 Hsp70 on different lysates with Rabbit anti-Hsp70 antibody (ET1601-11) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: A549 cell lysate
    Lane 3: MCF7 cell lysate
    Lane 4: HCT 116 cell lysate
    Lane 5: Mouse brain tissue lysate
    Lane 6: Mouse testis tissue lysate
    Lane 7: Rat testis tissue lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 70 kDa
    Observed band size: 70 kDa

    Exposure time: Lane 1-4: 43 seconds; Lane 5-7: 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 (ET1601-11) 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.

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

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