Anti-TSG101 抗体 [EPR7130(B)]

• IHC

Lab

Immunohistochemistry - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling TSG101 with ab125011 at a concentration of 0.1µg/ml.

The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5. ab125011 anti-TSG101 was incubated at 37°C for 16min.

Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling TSG101 with Purified ab125011 at 1 : 300 dilution (0.73 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

ab125011 staining TSG101 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab125011 at 1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid cancer tissue sections labeling TSG101 with Purified ab125011 at 1 : 300 dilution (0.73 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling TSG101 with Purified ab125011 at 1 : 50 dilution (4.5 µg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling TSG101 with Purified ab125011 at 1 : 20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling TSG101 with Purified ab125011 at 1 : 300 dilution (0.73 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling TSG101 with Purified ab125011 at 1 : 300 dilution (0.73 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• WB

Lab

Western blot - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

The doublet are consistent with what have been described in the literature (PMID : 27882925, 31684046, 32932791, 11427703).

All lanes:

Western blot - Anti-TSG101 antibody [EPR7130(B)] (ab125011) at 1/1000 dilution

Lane 1:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 2:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 6:

Mouse heart lysate at 20 µg

Lane 7:

Rat heart lysate at 20 µg

Lane 8:

Mouse brain lysate at 20 µg

Lane 9:

Rat brain lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 44 kDa

Observed band size: 47 kDa,52 kDa

false

• WB

Lab

Western blot - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

Different batches of ab125011 were tested on Mouse heart lysate at 1.0 μg/ml. 15 μg of lysate was loaded in each lane. Bands observed at 45 kDa.

This data was generarted using the unpurified format.

All lanes:

Western blot - Anti-TSG101 antibody [EPR7130(B)] (ab125011)

Predicted band size: 44 kDa

false