Antibodies, Proteins, Kits and Reagents for Life Science

348 441 ₸ Product size

100 µl 348 441 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Mouse monoclonal [4A10] to TSG101
Suitable for: Flow Cyt, WB, IHC-P
Reacts with: Mouse, Human
Isotype: IgG1

Product name

Anti-TSG101 antibody [4A10]
See all TSG101 primary antibodies
Description

Mouse monoclonal [4A10] to TSG101
Host species

Mouse
Specificity

This antibody recognizes the TSG-101 protein, the product of a recently identified tumor susceptibility gene the inactivation of which in mouse fibroblasts results in cell transformation and the ability of those cells to form tumors in nude mice.

Tested Applications & Species

Application	Species
Flow Cyt	Human
IHC-P	Human
WB	Mouse Human

See all applications and species data

Immunogen

Fusion protein corresponding to Human TSG101 aa 167-374. Expressed in E.coli
Database link: Q99816
Positive control
- WB: Neuro-2a, C8D30, NIH/3T3, RAW 264.7, C2C12 , HeLa, HepG2, A431, K562 and THP-1 whole cell lysate, mouse testis. ICC/IF: Amyloid peptide-treated mouse astrocytes IHC-P: human ovarian cancer tissue, human breast carcinoma tissue. Flow cyt: THP-1 cells
General notes

This product was changed from ascites to tissue culture supernatant on 12th February 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer

pH: 7.40
Constituent: 100% PBS

w/o preservative
Concentration information loading...
Purity

Protein G purified
Purification notes

Purified from tissue culture supernatant by Protein G chromatography to at least 95% homogeneity as determined by SDS-PAGE.
Clonality

Monoclonal
Clone number

4A10
Myeloma

NS1
Isotype

IgG1
Light chain type

kappa
Research areas

Western blot - Anti-TSG101 antibody [4A10] (ab83)

All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

Lane 1 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
Lane 2 : C8D30 whole cell lysate
Lane 3 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 4 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 5 : C2C12 (mouse myoblast cell line) whole cell lysate

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated anti-mouse IgG antibody at 1/500 dilution

Predicted band size: 43 kDa

10% SDS PAGE

Running conditions: 80V for 15min then 140V for 40min

Blocking: 5% non-fat milk in TBST at room temperature for 60min.

Washing conditions: 5 ml TBST, 4 x 5min

Transfer conditions: Semi-dry, 18 V, 60min (NC membrane)

Exposure system: Trident plus Western HRP Substrate
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [4A10] (ab83)

Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling TSG101 with ab83 at 1/100 dilution. Cytoplasmic staining is observed. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min. (Cuisinart Electric Pressure Cooker #EPC-1200, choose "high pressure"). Endogenous peroxidase blocking: 3% H₂O₂, RT, 30min. Blocking condition: 1.5% goat serum (dilute goat serum by 1xPBS), RT, 30min. Primary antibody incubation: 4°C overnight. Secondary antibody incubation: HRP Kit (Mouse IgG), 1:200, RT, 30min. Washing: PBS, 2 x 5 minutes. DAB detection.
Flow Cytometry - Anti-TSG101 antibody [4A10] (ab83) Image from Kahlert C et al., J Biol Chem. 2014;289(7):3869-75. Fig 1(C).; doi: 10.1074/jbc.C113.532267.

Panc-1 and Serum Exosomes were characterized by the exosome-specific expression of TSG101 by FACS analysis.
Exosomes were attached to 4-μm aldehyde/sulfate latex beads by mixing ∼30 μg of exosomes in a 100-μl volume of beads for 2 hours at room temperature. This suspension was diluted to 1 ml with PBS, and the reaction was stopped with 100 mm glycine and 2% BSA in PBS. Exosome-bound beads were washed in PBS/1% BSA, blocked with 10% BSA, and stained for FACS with anti-TSG101 (1:400, ab83). An Alexa Fluor® 488 conjugated anti-mouse IgG was used as the secondary antibody.
Flow Cytometry - Anti-TSG101 antibody [4A10] (ab83)

Flow Cytometry analysis of THP-1 cells labeling TSG101 with ab83 at 1/25 dilution (red). Unlabelled sample was used as a control (black). A Dylight 488-conjugated secondary antibody was used for FACS analysis.
Western blot - Anti-TSG101 antibody [4A10] (ab83)

All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

Lane 1 : Non-transfected (–) 293T whole cell extracts
Lane 2 : TSG101 shRNA transfected (+) 293T whole cell extracts

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated anti-mouse IgG antibody

Developed using the ECL technique.

Predicted band size: 43 kDa

10% SDS-PAGE
Western blot - Anti-TSG101 antibody [4A10] (ab83)

All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

Lane 1 : Mouse testis tissue extract
Lane 2 : Mouse pancreas tissue extract

Lysates/proteins at 50 µg per lane.

Secondary
All lanes : HRP-conjugated anti-mouse IgG antibody

Developed using the ECL technique.

Predicted band size: 43 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [4A10] (ab83)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling TSG101 with ab83 at 1/100 dilution. Cytoplasmic staining is observed. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
Western blot - Anti-TSG101 antibody [4A10] (ab83)

All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

Lane 1 : K562 whole cell extract
Lane 2 : THP-1 whole cell extract
Lane 3 : HL-60 whole cell extract

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated anti-mouse IgG antibody

Developed using the ECL technique.

Predicted band size: 43 kDa

10% SDS-PAGE
Western blot - Anti-TSG101 antibody [4A10] (ab83)

All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

Lane 1 : NIH-3T3 whole cell lysate/extract
Lane 2 : JC (mouse mammary adenocarcinoma) whole cell lysate/extract
Lane 3 : BCL-1 whole cell lysate/extract

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated anti-mouse IgG antibody

Predicted band size: 43 kDa

10% SDS-PAGE
Western blot - Anti-TSG101 antibody [4A10] (ab83)

All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1/500 dilution

Lane 1 : A431 whole cell lysate
Lane 2 : HeLa whole cell lysate
Lane 3 : HepG2 whole cell lysate
Lane 4 : K562 whole cell lysate
Lane 5 : THP-1 whole cell lysate

Lysates/proteins at 30 µg per lane.

Predicted band size: 43 kDa
Western blot - Anti-TSG101 antibody [4A10] (ab83)

All lanes : Anti-TSG101 antibody [4A10] (ab83) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : A-431 whole cell lysate (ab7909)
Lane 3 : Jurkat whole cell lysate (ab7899)
Lane 4 : HEK-293 whole cell lysate (ab7902)

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Donkey polyclonal to Mouse IgG (IRDyeTM 700DX) at 1/10000 dilution

Predicted band size: 43 kDa
Observed band size: 49 kDa
why is the actual band size different from the predicted?

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