Anti-BST2/Tetherin antibody [EPR23597-266]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-BST2/Tetherin antibody [EPR23597-266] (AB272169)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Mouse splenocyte cells labelling BST2/Tetherin with ab272169 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 splenocytes. 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 1000 dilution.

• Flow Cyt

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Flow Cytometry - Anti-BST2/Tetherin antibody [EPR23597-266] (AB272169)

Flow cytometric analysis of Mouse splenocyte cells labelling BST2/Tetherin with ab272169 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). A Goat anti rabbit IgG (AlexaFluor 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Cells were stained with rabbit IgG (Left) or ab272169 (Right). Then stained with anti-CD45R conjugated to Alexa Fluor® 647.

Gated on viable cells.

• IP

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Immunoprecipitation - Anti-BST2/Tetherin antibody [EPR23597-266] (AB272169)

BST2/Tetherin was immunoprecipitated from 0.35 mg Mouse spleen tissue lysate with ab272169 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272169 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 : ab272169 IP in Mouse spleen tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab272169 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 [EPR23597-266] (ab272169)

Predicted band size: 20 kDa

Observed band size: 28-40 kDa

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• WB

Lab

Western blot - Anti-BST2/Tetherin antibody [EPR23597-266] (AB272169)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

BST2 is type II transmembrane glycoprotein with a molecular mass of 28-40 KD, and forms cysteine-linked dimers at 64 KD which is consistent to the literature(PMID : 22520941; PMID : 19737401).

All lanes:

Western blot - Anti-BST2/Tetherin antibody [EPR23597-266] (ab272169) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse thymus tissue lysate at 20 µg

Lane 3:

Mouse lymph node tissue lysate at 20 µg

Lane 4:

P388D1 (mouse lymphoma monocyte; macrophage), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 20 kDa

Observed band size: 28-40 kDa,64 kDa

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