Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

This data was developed using ab302954, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SK-MEL-2(human skin malignant melanoma cell) cells labelling CITED1 with ab302954 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

This data was developed using ab302954, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling CITED1 with ab302954 at 1/500 (1.022 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : No staining on human kidney.The section was incubated with ab302954 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

This data was developed using ab302954, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-MEL-2(human skin malignant melanoma cell) cells labelling CITED1 with ab302954 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in SK-MEL-2 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (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 ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

This data was developed using ab302954, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-MEL-28(human malignant melanoma cell) cells labelling CITED1 with ab302954 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in SK-MEL-28 cell line. Low expression control : A431. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (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 ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

This data was developed using ab302954, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SK-MEL-28(human malignant melanoma cell) cells labelling CITED1 with ab302954 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

This data was developed using ab302954, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized B16-F10 (mouse skin melanoma cell) cells labelling CITED1 with ab302954 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

This data was developed using ab302954, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized B16-F10 (mouse skin melanoma cell) cells labelling CITED1 with ab302954 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in B16-F10 cell line. Low expression control : MEF (PMID : 24319640) Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8) ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (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 ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• WB

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Western blot - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

This data was developed using ab302954, the same antibody clone in a different buffer formulation.

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

Negative control : Lung, kidney (PMID : 8901575).

The molecular weight observed is consistent with what has been described in the literature (PMID : 16864582).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

Exposure time :

Lane 1, 2 : 59 seconds

Lane 3-8 : 103 seconds

All lanes:

Western blot - Anti-CITED1 antibody [EPR24375-81] (<a href='/en-us/products/primary-antibodies/cited1-antibody-epr24375-81-ab302954'>ab302954</a>) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Human kidney tissue lysate at 20 µg

Lane 4:

Mouse testis tissue lysate at 20 µg

Lane 5:

Mouse lung tissue lysate at 20 µg

Lane 6:

Mouse kidney tissue lysate at 20 µg

Lane 7:

Rat testis tissue lysate at 20 µg

Lane 8:

Rat lung tissue 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: 30 kDa

true

• WB

Supplier Data

Western blot - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

This data was developed using ab302954, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : A431 Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. Lane 3 was developed using a high sensitivity ECL substrate. Exposure time : 180 seconds

All lanes:

Western blot - Anti-CITED1 antibody [EPR24375-81] (<a href='/en-us/products/primary-antibodies/cited1-antibody-epr24375-81-ab302954'>ab302954</a>) at 1/1000 dilution

Lane 1:

SK-MEL-2 (human skin malignant melanoma cell) whole cell lysate at 20 µg

Lane 2:

SK-MEL-28 (human malignant melanoma cell) whole cell lysate at 20 µg

Lane 3:

A431 (human epidermoid carcinoma epithelial cell) 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: 30 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CITED1 antibody [EPR24375-81] - BSA and Azide free (AB302955)

This data was developed using ab302954, the same antibody clone in a different buffer formulation.

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

Low expression : MEF (PMID : 22958931).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-CITED1 antibody [EPR24375-81] (<a href='/en-us/products/primary-antibodies/cited1-antibody-epr24375-81-ab302954'>ab302954</a>) at 1/1000 dilution

Lane 1:

B16-F10 (mouse skin melanoma cell), whole cell lysate at 20 µg

Lane 2:

MEF (mouse embryo fibroblast), 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: 30 kDa

true

Exposure time: 3min