Anti-CITED1 antibody [EPR24375-81]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CITED1 antibody [EPR24375-81] (AB302954)

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] (AB302954)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling CITED1 with ab302954 at 1/500 (1.022 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on human testis. 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED1 antibody [EPR24375-81] (AB302954)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling CITED1 with ab302954 at 1/500 (1.022 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. 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] (AB302954)

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

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Immunocytochemistry/ Immunofluorescence - Anti-CITED1 antibody [EPR24375-81] (AB302954)

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)

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Flow Cytometry (Intracellular) - Anti-CITED1 antibody [EPR24375-81] (AB302954)

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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED1 antibody [EPR24375-81] (AB302954)

Immunohistochemical analysis of paraffin-embedded Human thyroid cancer tissue labeling CITED1 with ab302954 at 1/500 (1.022 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on human thyroid cancer (PMID : 15272279).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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED1 antibody [EPR24375-81] (AB302954)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling CITED1 with ab302954 at 1/500 (1.022 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on mouse testis. 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED1 antibody [EPR24375-81] (AB302954)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling CITED1 with ab302954 at 1/500 (1.022 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on rat testis.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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CITED1 antibody [EPR24375-81] (AB302954)

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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CITED1 antibody [EPR24375-81] (AB302954)

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] (AB302954)

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] (ab302954) 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] (AB302954)

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] (ab302954) 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] (AB302954)

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] (ab302954) 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