Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free
See all NSUN2/SAKI primary antibodies
Description

Rabbit monoclonal [EPR24140-93] to NSUN2/SAKI - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HepG2, HeLa, HEK-293 and A431 whole cell lysates; ES-D3 whle cell lysate; Mouse spleen tissue lysate; Rat cerebellum tissue lysate. IHC-P: Human pancreas and colon carcinoma tissue; Mouse skin tissue; Rat colon tissue. ICC: HeLa and NIH/3T3 cells. Flow cyt: HeLa and ES-D3 cells. IP: HEK-293 whole cell lysate; Mouse spleen tissue lysate.
General notes
ab277534 is the carrier-free version of ab259941. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab277534 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C.
Storage buffer

Constituent: 100% PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR24140-93
Isotype

IgG
Research areas

Western blot - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

All lanes : Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941) at 1/1000 dilution

Lane 1 : HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
Lane 4 : A431 (human epidermoid carcinoma epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

Predicted band size: 86 kDa
Observed band size: 100 kDa
why is the actual band size different from the predicted?

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

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

The observed MW is consistent with what has described in the literature (PMID: 23604283, 25233213, 27447970, 31487418).

Exposure times: Lane 1: 15 seconds

Lane 2-4: 48 seconds.
Immunocytochemistry - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling NSUN2/SAKI with ab259941 at 1/1000 dilution (0.503 μg/ml), followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) antibody at 1/1000 dilution (Green). Confocal image showing weak cytoplasmic and strong nuclear staining in HeLa cells. 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling NSUN2/SAKI with ab259941 at 1/200 dilution (2.515 μg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Nuclear staining on human pancreas (PMID: 16713953). The section was incubated with ab259941 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).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunoprecipitation - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

NSUN2/SAKI was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate with ab259941 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259941 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10 ug

Lane 2: ab259941 IP in HEK-293 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259941 in HEK-293 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 15 seconds.
Western blot - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

All lanes : Anti-NSUN2/SAKI antibody [EPR24140-93] (ab259941) at 1/1000 dilution

Lane 1 : ES-D3 (mouse embryonic mtipotent stem Cell) whole cell lysate
Lane 2 : Mouse spleen tissue lysate
Lane 3 : Rat cerebellum tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

Predicted band size: 86 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

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

The observed MW is consistent with what has described in the literature (PMID: 27306184).

Exposure times: Lane 1: 48 seconds

Lane 2, 3: 3 minutes.
Immunocytochemistry - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) cells labelling NSUN2/SAKI with ab259941 at 1/1000 (0.503 µg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) antibody at 1/1000 dilution (Green). Confocal image showing weak cytoplasmic and strong nuclear staining in NIH/3T3 cells. 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling NSUN2/SAKI with ab259941 at 1/200 dilution (2.515 μg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Nuclear staining on mouse skin (PMID: 16713953). The section was incubated with ab259941 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).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunoprecipitation - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

NSUN2/SAKI was immunoprecipitated from 0.35 mg mouse spleen tissue lysate with ab259941 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259941 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse spleen tissue lysate 10 ug

Lane 2: ab259941 IP in Mouse spleen tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259941 in mouse spleen tissue lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 15 seconds.
Flow Cytometry - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling NSUN2/SAKI with ab259941 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labeling NSUN2/SAKI with ab259941 at 1/200 dilution (2.515 μg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Nuclear staining on human colon carcinoma (PMID: 16713953). The section was incubated with ab259941 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).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Flow Cytometry - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized ES-D3 (mouse embryonic multipotent stem cell) cells labelling NSUN2/SAKI with ab259941 at 1/50 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)

This data was developed using ab259941, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling NSUN2/SAKI with ab259941 at 1/200 dilution (2.515 µg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Nuclear staining on rat colon.The section was incubated with ab259941 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).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Anti-NSUN2/SAKI antibody [EPR24140-93] - BSA and Azide free (ab277534)