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TSG101 Recombinant Rabbit Monoclonal Antibody [JJ0900]

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Specification

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Catalog# ET1701-59

TSG101 Recombinant Rabbit Monoclonal Antibody [JJ0900]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

TSG101 Recombinant Rabbit Monoclonal Antibody [JJ0900]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human TSG101 aa 24-73 / 390.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 44 kDa

Positive Control

HeLa cell lysate, K-562 cell lysate, MCF7 cell lysate, Jurkat cell lysate, C2C12 cell lysate, PC-12 cell lysate, NIH/3T3 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, HeLa, NIH/3T3, PC-12, human kidney tissue, mouse colon tissue, mouse kidney tissue, human colon carcinoma tissue.

Conjugation

unconjugated

Clone Number

JJ0900

RRID

AB_3068599

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:200

  • IF-Tissue

  • 1:200-1:500

  • IHC-P

  • 1:500-1:1,000

  • FC

  • 1:1,000

Target

Function

The transformation of a normal cell to one that is malignant can result from mutations in genes that encode proteins with key regulatory functions. Examples include the retinoblastoma gene product (Rb p110), p53, VHL and APC. Using a novel cloning strategy that allows the isolation of previously uncharacterized genes encoding selectable recessive phenotypes, an additional tumor suppressor gene has been identified. This gene, termed tsg 101 for tumor susceptibility gene 101, encodes a stathmin binding domain protein. When expression of this growth inhibitory gene is blocked in NIH/3T3 cells using antisense mRNA, the cells exhibit a transformed phenotype and are tumorigenic in SL6 mice.

Background References

1. Ruiz-Guillen M et al. Capsid-deficient alphaviruses generate propagative infectious microvesicles at the plasma membrane. Cell Mol Life Sci 73:3897-916 (2016).

2. Baranyai T et al. Isolation of Exosomes from Blood Plasma: Qualitative and Quantitative Comparison of Ultracentrifugation and Size Exclusion Chromatography Methods. PLoS One 10:e0145686 (2015).

Sequence Similarity

Belongs to the ubiquitin-conjugating enzyme family. UEV subfamily.

Tissue Specificity

Heart, brain, placenta, lung, liver, skeletal, kidney and pancreas.

Post-translational Modification

Monoubiquitinated at multiple sites by LRSAM1 and by MGRN1. Ubiquitination inactivates it, possibly by regulating its shuttling between an active membrane-bound protein and an inactive soluble form. Ubiquitination by MGRN1 requires the presence of UBE2D1.

Subcellular Location

Cytoplasm, Cytoskeleton, Endosome, Membrane, Nucleus.

UNIPROT

SwissProt: Q99816 Human

SwissProt: Q61187 Mouse

SwissProt: Q6IRE4 Rat

Synonyms

ESCRT I complex subunit TSG101 antibody

ESCRT-I complex subunit TSG101 antibody

TS101_HUMAN antibody

TSG 10 antibody

TSG 101 antibody

TSG10 antibody

Tsg101 antibody

Tumor susceptibility gene 10 antibody

Tumor susceptibility gene 101 antibody

Tumor susceptibility gene 101 protein antibody

Expand

Images

  • 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<br />Lane 2: K-562 cell lysate<br />Lane 3: MCF7 cell lysate<br />Lane 4: Jurkat cell lysate<br />Lane 5: C2C12 cell lysate<br />Lane 6: PC-12 cell lysate<br /><br />Lysates/proteins at 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; 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/ET1701-59" 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
    Lane 2: K-562 cell lysate
    Lane 3: MCF7 cell lysate
    Lane 4: Jurkat cell lysate
    Lane 5: C2C12 cell lysate
    Lane 6: PC-12 cell lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 3 minutes 20 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 (ET1701-59) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of TSG101 with anti-TSG101 antibody[JJ0900] (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/2,000 dilution.<br />Lane 1: Wild-type Hela whole cell lysate (10 µg).<br />Lane 2/3: TSG101 knockdown Hela whole cell lysate (10 µg).<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in 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>, 1/2,000) 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 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of TSG101 with anti-TSG101 antibody[JJ0900] (ET1701-59) at 1/2,000 dilution.
    Lane 1: Wild-type Hela whole cell lysate (10 µg).
    Lane 2/3: TSG101 knockdown Hela whole cell lysate (10 µg).

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1701-59, 1/2,000) was used in 5% BSA 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 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.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: K-562 cell lysate<br />Lane 3: Jurkat cell lysate<br />Lane 4: NIH/3T3 cell lysate<br />Lane 5: PC-12 cell lysate<br />Lane 6: Mouse brain tissue lysate<br />Lane 7: Rat brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 44 kDa<br />Observed band size: 44/47 kDa<br /><br />Exposure time: 1 minute 40 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/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</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 TSG101 on different lysates with Rabbit anti-TSG101 antibody (ET1701-59) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: K-562 cell lysate
    Lane 3: Jurkat cell lysate
    Lane 4: NIH/3T3 cell lysate
    Lane 5: PC-12 cell lysate
    Lane 6: Mouse brain tissue lysate
    Lane 7: Rat brain tissue lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 1 minute 40 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 (ET1701-59) 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.

  • Immunocytochemistry analysis of HeLa cells labeling TSG101 with Rabbit anti-TSG101 antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/200 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-TSG101 antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/200 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. 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 TSG101 with Rabbit anti-TSG101 antibody (ET1701-59) at 1/200 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-TSG101 antibody (ET1701-59) at 1/200 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.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling TSG101 with Rabbit anti-TSG101 antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/200 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-TSG101 antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/200 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. 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 NIH/3T3 cells labeling TSG101 with Rabbit anti-TSG101 antibody (ET1701-59) at 1/200 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-TSG101 antibody (ET1701-59) at 1/200 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.

  • Immunocytochemistry analysis of PC-12 cells labeling TSG101 with Rabbit anti-TSG101 antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/200 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-TSG101 antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/200 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. 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 PC-12 cells labeling TSG101 with Rabbit anti-TSG101 antibody (ET1701-59) at 1/200 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-TSG101 antibody (ET1701-59) at 1/200 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.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-TSG101 antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-TSG101 antibody (ET1701-59) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-59) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-TSG101 antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-TSG101 antibody (ET1701-59) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-59) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of HeLa cells labeling TSG101.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1701-59" style="font-weight: bold;text-decoration: underline;">ET1701-59</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling TSG101.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-59, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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