Anti-BST2/Tetherin antibody [EPR20202-150] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-BST2/Tetherin antibody [EPR20202-150] - BSA and Azide free (AB243561)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling BST2/Tetherin with ab243230 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody (Green). Confocal image showing cytoplasmic staining in HeLa cells (PMID : 20529266).

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243230).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-BST2/Tetherin antibody [EPR20202-150] - BSA and Azide free (AB243561)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-937 (human histiocytic lymphoma cell line) cells labeling BST2/Tetherin with ab243230 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody (Green). Confocal image showing cytoplasmic staining in U-937 cells (PMID : 20529266).

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243230).

• Flow Cyt

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Flow Cytometry - Anti-BST2/Tetherin antibody [EPR20202-150] - BSA and Azide free (AB243561)

Flow cytometric analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling BST2/Tetherin with ab243230 at 1/700 dilution (Red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243230).

• IP

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Immunoprecipitation - Anti-BST2/Tetherin antibody [EPR20202-150] - BSA and Azide free (AB243561)

BST2/Tetherin was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab243230 at 1/40 dilution. Western blot was perfromed on the immunoprecipitate using ab243230 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

Lane 1 : HeLa whole cell lysate 10 μg (input)
Lane 2 : ab243230 IP in HeLa whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab243230 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST
Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243230).

All lanes:

Immunoprecipitation - Anti-BST2/Tetherin antibody [EPR20202-150] (<a href='/en-us/products/primary-antibodies/bst2-tetherin-antibody-epr20202-150-ab243230'>ab243230</a>)

Predicted band size: 20 kDa

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• WB

Lab

Western blot - Anti-BST2/Tetherin antibody [EPR20202-150] - BSA and Azide free (AB243561)

This data was developed using the same antibody clone in a different buffer formulation (ab243230).

Lanes 1 - 4 : Merged signal (red and green). Green - ab243230 observed at 30 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab243230 was shown to react with BST2/Tetherin in wild-type HeLa cells in Western blot with loss of signal observed in BST2 knockout samples. Wild-type HeLa and BST2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab243230 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-BST2/Tetherin antibody [EPR20202-150] (<a href='/en-us/products/primary-antibodies/bst2-tetherin-antibody-epr20202-150-ab243230'>ab243230</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Bst2 knockout HeLa cell lysate at 20 µg

Lane 3:

Wild-type A431 cell lysate at 20 µg

Lane 4:

BST2 knockout A431 cell lysate at 20 µg

Predicted band size: 20 kDa

Observed band size: 30 kDa

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