Anti-Myc tag antibody [Hyper-myc] - Mouse IgG2a (Chimeric)

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Myc tag antibody [Hyper-myc] - Mouse IgG2a (Chimeric) (AB289980)

Intracellular flow cytometric analysis of 293T (human embryonic kidney) transfected with TMEM-119 expression vector containing a myc-His-tag®, fixed with 2% paraformaldehyde and permeabelized 0.1% Tween-20. Cells were surface stained with TMEM-119 conjugated with Alexa Fluor® 647 (ab225494). Then fixed with 2% PFA for 10min followed by intracellularly stained with mouse IgG or ab289980. ab289980 was used at 1 : 1000 dilution (0.1μg)(Right) compared with Mouse monoclonal IgG (Left). Secondary antibody used was Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution.

• ICC

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Immunocytochemistry - Anti-Myc tag antibody [Hyper-myc] - Mouse IgG2a (Chimeric) (AB289980)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T cells transfected with CRISPR-Cas9 (S. Aureas) expression vector containing a myc tag, labeling myc-tag with ab289980 at 1/500 (2.202 μg/ml) dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) (ab150117) preadsorbed secondary antibody at 1/1000 dilution (Green). Confocal image showing positive staining in HEK-293T cells transfected with CRISPR-Cas9 (S. Aureas) expression vector containing a myc tag.

Nulcear counterstain is DAPI. Cells were also counterstained with ab203933 Anti-CRISPR-Cas9 rabbit monoclonal antibody at 1/500 (2 μg/ml) dilution, followed by secondary antibody ab203933 Anti-CRISPR-Cas9 rabbit monoclonal antibody at 1/200 (2.5 μg/ml) dilution.

-ve control 1 : ab289980 at 1/500 (2.202 μg/ml) dilution, followed by ab15088 1/200 (2.5 μg/ml)

-ve control 2 : ab15088 1/200 (2 μg/ml), followed by ab150117 1/1000 (2 μg/ml)

• IP

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Immunoprecipitation - Anti-Myc tag antibody [Hyper-myc] - Mouse IgG2a (Chimeric) (AB289980)

Myc-tag was immunoprecipitated from HEK-293T transfected with human NF2 S518A expression vector containing a myc-His-tag® whole cell lysate (2μg). Western blot was performed using immunopreciptate, ab289980 was used at 1/30 dilution (2μg in 0.35mg lysates). Secondary antibody used was Veriblot for IP secondary antibody (HRP) (ab131366) at 1/5000 dilution.

Lane 1 (Input) : HEK-293T transfected with human NF2 S518A expression vector containing a myc-His-tag® whole cell lysate, 2μg.

Lane 2 (+) : HEK-293T transfected with human NF2 S518A expression vector containing a myc-His-tag® whole cell lysate

Lane 3 (-) : Human IgG instead of ab289980 in HEK-293T transfected with human NF2 S518A expression vector containing a myc-His-tag® whole cell lysate

Blocking and dilution buffer : 5% NFDM/TBST.

ab289980 detects a ranged of myc tagged proteins in WB.

Lane 1:

Immunoprecipitation - Anti-Myc tag antibody [Hyper-myc] - Mouse IgG2a (Chimeric) (ab289980)

Lane 1:

Immunoprecipitation - Anti-Myc tag antibody [Hyper-myc] (<a href='/en-us/products/primary-antibodies/myc-tag-antibody-hyper-myc-ab289981'>ab289981</a>)

Predicted band size: 49 kDa

Observed band size: 70 kDa

false

Exposure time: 6s

• ELISA

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ELISA - Anti-Myc tag antibody [Hyper-myc] - Mouse IgG2a (Chimeric) (AB289980)

Indirect ELISA antibody dose-response curve of ab289980 for Human myc-tag antigen, at a concentration of 1000ng/ml. Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Mouse IgG (H+L) at 1/1000 dilution.

• WB

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Western blot - Anti-Myc tag antibody [Hyper-myc] - Mouse IgG2a (Chimeric) (AB289980)

Blocking and dilution buffer : 5% NFDM/TBST

ab289980 detects a ranged of myc tagged proteins in WB.

All lanes:

Western blot - Anti-Myc tag antibody [Hyper-myc] - Mouse IgG2a (Chimeric) (ab289980) at 1/5000 dilution

Lane 1:

HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate at 1 µg

Lane 2:

HEK-293T transfected with cas9 ( S. Aureas) expression vector containing a myc-His-tag®, whole cell lysate at 1 µg

Lane 3:

HEK-293T transfected with human SNCA140 expression vector containing a myc-His-tag®, whole cell lysate at 1 µg

Lane 4:

HEK-293T transfected with human IDS expression vector containing a myc-His-tag®, whole cell lysate at 1 µg

Lane 5:

HEK-293T transfected with human NF2 S518A expression vector containing a myc-His-tag®, whole cell lysate at 1 µg

Lane 6:

HEK-293T transfected with human Alphasynuclein expression vector containing a myc-His-tag®, whole cell lysate at 1 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG Fc (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-fc-hrp-preadsorbed-ab98717'>ab98717</a>) at 1/10000 dilution

Predicted band size: 49 kDa

false

Exposure time: 3s