Anti-BTN3A1 antibody [EPR28070-15]

• IP

Supplier Data

Immunoprecipitation - Anti-BTN3A1 antibody [EPR28070-15] (AB315804)

BTN3A1 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate with ab315804 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315804 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-BTN3A1 antibody [EPR28070-15] (ab315804) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate

Lane 2:

ab315804 at 1/30 IP in Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab315804 in Jurkat whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-BTN3A1 antibody [EPR28070-15] (AB315804)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

BTN3A1 expression can be induced by PMA treatment (PMID : 27911820).

The identity of the higher MW band at approximately 200 kDa in lane 1 is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-BTN3A1 antibody [EPR28070-15] (ab315804) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 4:

Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 5:

THP-1 treated with 80nM Phorbol-12-myristate-13-acetate (PMA) for 72 hours, whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 58 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-BTN3A1 antibody [EPR28070-15] (AB315804)

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

Low expression : hypothalamus, brain (PMID : 9149941).

The identity of the higher MW band at approximately 200 kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-BTN3A1 antibody [EPR28070-15] (ab315804) at 1/1000 dilution

Lane 1:

Human placenta tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Human hypothalamus tissue lysate at 20 µg

Lane 4:

Human brain tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 58 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-BTN3A1 antibody [EPR28070-15] (AB315804)

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

This antibody does not cross-react with human BTN3A2 and BTN3A3.

All the recombinant proteins were made in-house.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-BTN3A1 antibody [EPR28070-15] (ab315804) at 1/1000 dilution

Lane 1:

His-tagged human BTN3A1 recombinant protein fragment at 10 ng

Lane 2:

His-tagged human BTN3A2 recombinant protein at 10 ng

Lane 3:

His-tagged human BTN3A3 Synuclein recombinant protein at 10 ng

Secondary

All lanes:

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

Observed band size: 28 kDa

false

Exposure time: 37s