Anti-Human Nucleoli antibody [NM95] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Human Nucleoli antibody [NM95] - BSA and Azide free (AB234031)

This data was developed using ab190710, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol, 0.1% TritonX-100 permeabilized K562 (Human chronic myelogenous leukemia lymphoblast cell line ) cells labelling Human Nucleoli with ab190710 at 1/50 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) (ab150113) secondary antibody at 1/1000 dilution (green). Confocal image showing nucleolar staining in K562 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution (red).

Tubulin is detected with Anti-beta Tubulin rabbit monoclonal antibody (ab179513) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) (ab150080) secondary antibody at 1/500 dilution (red).

The negative controls are as follows :
-ve control 1 : ab190710 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) (ab150080) secondary at 1/1000 dilution.
-ve control 2 : Anti-beta Tubulin rabbit monoclonal antibody (ab179513) at 1/200 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) (ab150113) secondary at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Human Nucleoli antibody [NM95] - BSA and Azide free (AB234031)

This data was developed using ab190710, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol, 0.1% TritonX-100 permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Human Nucleoli with ab190710 at 1/50 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) (ab150113) secondary antibody at 1/1000 dilution (green). Confocal image showing nucleolar staining in HeLa cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-beta Tubulin rabbit monoclonal antibody (ab179513) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) (ab150080) secondary antibody at 1/500 dilution (red).

The negative controls are as follows :
-ve control 1 : ab190710 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) (ab150080) secondary at 1/500 dilution.
-ve control 2 : Anti-beta Tubulin rabbit monoclonal antibody (ab179513) at 1/200 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) (ab150113) secondary at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Human Nucleoli antibody [NM95] - BSA and Azide free (AB234031)

This data was developed using ab190710, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded sections human colon cancer tissue labelling Human Nucleoli with ab190710 at 1/2000 dilution, followed by secondary ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879). Nuclear staining in human colon cancer tissue is observed. Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879).

The section was incubated with ab190710 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Human Nucleoli antibody [NM95] - BSA and Azide free (AB234031)

This data was developed using ab190710, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human kidney tissue labelling Human Nucleoli with ab190710 at 1/1000 dilution, followed by secondary antibody the ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879). Nuclear staining in human kidney is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

The section was incubated with ab190710 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Human Nucleoli antibody [NM95] - BSA and Azide free (AB234031)

This data was developed using ab190710, the same antibody clone in a different buffer formulation.

Flow cytometry overlay histogram of 4% paraformaldehyde-fixed, 90% methanol permeabilized K562 (Human chronic myelogenous leukemia lymphoblast) cells labelling Human Nucleoli with ab190710 at 1/1000 dilution (red) compared with a Mouse monoclonal IgG (black) and an unlabelled control (Cell without incubation with primary antibody and secondary antibody) (blue). Goat anti mouse IgG (Alexa Fluor^® 488) (ab150113), at 1/2000 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-Human Nucleoli antibody [NM95] - BSA and Azide free (AB234031)

This data was developed using ab190710, the same antibody clone in a different buffer formulation.

Exposure times :
lane 1 : 3 seconds
lane 2 : 5 seconds

5% NFDM/TBST

Lysates were made freshly and used in WB immediately to minimize protein degradation.

All lanes:

Western blot - Anti-Human Nucleoli antibody [NM95] - Nucleolar Marker (<a href='/en-us/products/primary-antibodies/human-nucleoli-antibody-nm95-nucleolar-marker-ab190710'>ab190710</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 100 kDa

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