Anti-UPF3B/RENT3B + UPF3A antibody [EPR24112-81] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-UPF3B/RENT3B + UPF3A antibody [EPR24112-81] - BSA and Azide free (AB288302)

This data was developed using ab269998, 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 UPF3A with ab269998 at 1/50 dilution (0.1 μg) (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

Lab

Immunocytochemistry/ Immunofluorescence - Anti-UPF3B/RENT3B + UPF3A antibody [EPR24112-81] - BSA and Azide free (AB288302)

This data was developed using ab269998, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling UPF3A with ab269998 at 1/100 (5.35 μg/ml) 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 HeLa cell line. 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 : ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• IP

Lab

Immunoprecipitation - Anti-UPF3B/RENT3B + UPF3A antibody [EPR24112-81] - BSA and Azide free (AB288302)

This data was developed using ab269998, the same antibody clone in a different buffer formulation.

UPF3A was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 μg with ab269998 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab269998 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : (Input) HeLa whole cell lysate 10 μg

Lane 2 : HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab269998 in HeLa whole cell lysate

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-UPF3B/RENT3B + UPF3A antibody [EPR24112-81] (<a href='/en-us/products/primary-antibodies/upf3b-rent3b-upf3a-antibody-epr24112-81-ab269998'>ab269998</a>)

Observed band size: 60 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-UPF3B/RENT3B + UPF3A antibody [EPR24112-81] - BSA and Azide free (AB288302)

This data was developed using ab269998, the same antibody clone in a different buffer formulation.

5% NFDM/TBST was used as a blocking and dilution buffer.

This antibody (ab269998) reacts with an unidentifiable protein around 35 kDa.

The zebrafish embryo tissue lysate is kindly provided by Prof. Jun Chen, Zhejiang University.

All lanes:

Western blot - Anti-UPF3B/RENT3B + UPF3A antibody [EPR24112-81] (<a href='/en-us/products/primary-antibodies/upf3b-rent3b-upf3a-antibody-epr24112-81-ab269998'>ab269998</a>) at 1/1000 dilution

Lane 1:

Zebrafish embryo tissue lysate at 10 µg

Lane 2:

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

Lane 3:

293T (human embryonic kidney 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: 60 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-UPF3B/RENT3B + UPF3A antibody [EPR24112-81] - BSA and Azide free (AB288302)

This data was developed using ab269998, the same antibody clone in a different buffer formulation.

5% NFDM/TBST was used as a blocking and diliution buffer.

Exposure time : 3.25 seconds.

This antibody has cross-reaction with zebrafish Upf3b.
Both samples are recombinant proteins expressed from an E.coli expression system.

All lanes:

Western blot - Anti-UPF3B/RENT3B + UPF3A antibody [EPR24112-81] (<a href='/en-us/products/primary-antibodies/upf3b-rent3b-upf3a-antibody-epr24112-81-ab269998'>ab269998</a>) at 1/1000 dilution

Lane 1:

Truncated Zebrafish Upf3a recombinant protein (aa32-261, predicted MW 28 kDa), 10 ng

Lane 2:

Full-length Zebrafish Upf3b recombinant protein (aa1-467) 10 ng

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: 60 kDa

false