重组Anti-Ras抗体[EPR3255]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Ras antibody [EPR3255] (AB108602)

Intracellular Flow Cytometry analysis of HEK-293 (Human embryonic kidney epithelial cell) cells labeling Ras with purified ab108602 at 1/30 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Ras antibody [EPR3255] (AB108602)

Immunocytochemistry/ Immunofluorescence analysis of 293T (Human embryonic kidney epithelial cell) cells labeling Ras with purified ab108602 at 1 : 100 dilution (10 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras antibody [EPR3255] (AB108602)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labelling Ras with ab209974 at 1/10000 (0.104 μg/ml) dilution followed by a LeicaDS9800 (Bond^™ Polymer Refine Detection) at ready to use dilution. Positive staining on human breast carcinoma. The section was incubated with ab209974 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 LeicaDS9800 (Bond^™ Polymer Refine Detection) at ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. This data was developed using the same antibody clone in a different buffer formulation (ab209974).

• IP

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Immunoprecipitation - Anti-Ras antibody [EPR3255] (AB108602)

ab108602 (purified) at 1/20 dilution (2 µg) immunoprecipitating Ras in SH-SY5Y whole cell lysate.
Lane 1 (input) : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+) : ab108602 & SH-SY5Y whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab108602 in SH-SY5Y whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Ras antibody [EPR3255] (ab108602)

Predicted band size: 21 kDa

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• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras antibody [EPR3255] (AB108602)

Immunohistochemical analysis of paraffin-embedded rat cardiac muscle tissue labelling Ras with ab209974 at 1/10000 (0.104 μg/ml) dilution followed by a LeicaDS9800 (Bond^™ Polymer Refine Detection) at ready to use dilution. Positive staining on rat cardiac muscle. The section was incubated with ab209974 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 LeicaDS9800 (Bond^™ Polymer Refine Detection) at ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. This data was developed using the same antibody clone in a different buffer formulation (ab209974).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras antibody [EPR3255] (AB108602)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labelling Ras with ab209974 at 1/10000 (0.104 μg/ml) dilution followed by a LeicaDS9800 (Bond^™ Polymer Refine Detection) at ready to use dilution. Positive staining on mouse cerebrum. The section was incubated with ab209974 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 LeicaDS9800 (Bond^™ Polymer Refine Detection) at ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. This data was developed using the same antibody clone in a different buffer formulation (ab209974).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ras antibody [EPR3255] (AB108602)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labelling Ras with ab209974 at 1/10000 (0.104 μg/ml) dilution followed by a LeicaDS9800 (Bond^™ Polymer Refine Detection) at ready to use dilution. Positive staining on rat cerebrum. The section was incubated with ab209974 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 LeicaDS9800 (Bond^™ Polymer Refine Detection) at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. This data was developed using the same antibody clone in a different buffer formulation (ab209974).

• WB

Lab

Western blot - Anti-Ras antibody [EPR3255] (AB108602)

All lanes:

Western blot - Anti-Ras antibody [EPR3255] (ab108602) at 1/10000 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

Lane 2:

Rat brain lysates at 20 µg

Lane 3:

Mouse heart lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

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• WB

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Western blot - Anti-Ras antibody [EPR3255] (AB108602)

All lanes:

Western blot - Anti-Ras antibody [EPR3255] (ab108602) at 1/1000 dilution

Lane 1:

HEK-293 whole cell lysate at 10 µg

Lane 2:

SH-SY5Y cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 21 kDa

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• WB

Lab

Western blot - Anti-Ras antibody [EPR3255] (AB108602)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of rat brain lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab108602 (1/10,000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/20,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique. ab108602 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant for temperatures ranging between 4°C, 22°C and 37°C, demonstrating that antibody activity is not affected

All lanes:

Western blot - Anti-Ras antibody [EPR3255] (ab108602) at 1/10000 dilution

All lanes:

Rat brain lysate at 15 µg with NDFM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

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Exposure time: 5s