Anti-BST2/Tetherin antibody [EPR23597-202]

• Flow Cyt

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Flow Cytometry - Anti-BST2/Tetherin antibody [EPR23597-202] (AB246508)

Flow cytometric analysis of Mouse splenocyte cells labelling BST2/Tetherin with ab246508 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left).A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Cells were stained with rabbit IgG (Left) or ab246508 (Right). Then stained with anti-CD45R conjugated to Alexa Fluor® 647.

Gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BST2/Tetherin antibody [EPR23597-202] (AB246508)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling BST2/Tetherin with ab246508 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse spleen (PMID : 19903902).The section was incubated with ab246508 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-BST2/Tetherin antibody [EPR23597-202] (AB246508)

Immunofluorescent analysis of 100% methanol-fixed EL.4 cells labelling BST2/Tetherin with ab246508 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in EL.4 cell line. 100% methanol fixation is recommended. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BST2/Tetherin antibody [EPR23597-202] (AB246508)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling BST2/Tetherin with ab246508 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : No staining on mouse skeletal muscle (PMID : 19903902). The section was incubated with ab246508 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-BST2/Tetherin antibody [EPR23597-202] (AB246508)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Mouse splenocyte cells labelling BST2/Tetherin with ab246508 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplamic staining in mouse splenocyte ab195889 Anti-alpha Tubulin antibody (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IP

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Immunoprecipitation - Anti-BST2/Tetherin antibody [EPR23597-202] (AB246508)

BST2/Tetherin was immunoprecipitated from 0.35 mg Mouse spleen tissue lysate 10ug with ab246508 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246508 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse spleen tissue lysate 10ug

Lane 2 : ab246508 IP in Mouse spleentissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab246508 in mouse spleen tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 15 seconds

BST2 is type II transmembrane glycoprotein with a molecular mass of 28-40 KD, which is consistent to the literature(PMID : 22520941; PMID : 19737401).

All lanes:

Immunoprecipitation - Anti-BST2/Tetherin antibody [EPR23597-202] (ab246508)

Predicted band size: 20 kDa

Observed band size: 28-40 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-BST2/Tetherin antibody [EPR23597-202] (AB246508)

BST2/Tetherin was immunoprecipitated from 0.35 mg EL4 (mouse lymphoma T lymphocyte) whole cell lysate with ab246508 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246508 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : EL4 (mouse lymphoma T lymphocyte), whole cell lysate 10 ug

Lane 2 : ab246508 IP in EL4 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab246508 in EL4 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 15 seconds

BST2 is type II transmembrane glycoprotein with a molecular mass of 28-40 KD, which is consistent to the literature(PMID : 22520941; PMID : 19737401).

All lanes:

Immunoprecipitation - Anti-BST2/Tetherin antibody [EPR23597-202] (ab246508)

Predicted band size: 20 kDa

Observed band size: 28-40 kDa

false