Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] - BSA and Azide free (AB251321)

This data was developed using ab200829, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling NFIB / NF1B2 + NFIC with ab200829 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human breast carcinoma tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] - BSA and Azide free (AB251321)

This data was developed using ab200829, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling NFIB / NF1B2 + NFIC with ab200829 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mouse cardiac muscle tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] - BSA and Azide free (AB251321)

This data was developed using ab200829, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Based on the sequence analysis, ab200829 recognizes six isoforms within human with predicted MW's of 56KDa, 55KDa, 48KDa, 45KDa, 48KDa and 49KDa, respectively.

All lanes:

Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (<a href='/en-us/products/primary-antibodies/nfib-nf1b2-nfic-ctf-antibody-epr14504-ab200829'>ab200829</a>) at 1/10000 dilution

All lanes:

HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 47 kDa,56 kDa

Observed band size: 45-65 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] - BSA and Azide free (AB251321)

This data was developed using ab200829, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Based on the sequence analysis, ab200829 recognizes six isoforms within human with predicted Mw’s of 56KDa, 55KDa, 48KDa, 45KDa, 48KDa and 49KDa, respectively.

All lanes:

Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (<a href='/en-us/products/primary-antibodies/nfib-nf1b2-nfic-ctf-antibody-epr14504-ab200829'>ab200829</a>) at 1/10000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 47 kDa,56 kDa

Observed band size: 45-65 kDa

false

Exposure time: 30s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] - BSA and Azide free (AB251321)

This data was developed using ab200829, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling NFIB / NF1B2 + NFIC with ab200829 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on HeLa cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows : -
-ve control 1 : ab200829 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] - BSA and Azide free (AB251321)

This data was developed using ab200829, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human cervix carcinoma tissue labeling NFIB / NF1B2 + NFIC with ab200829 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human cervix carcinoma tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] - BSA and Azide free (AB251321)

This data was developed using ab200829, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling NFIB / NF1B2 + NFIC with ab200829 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] - BSA and Azide free (AB251321)

This data was developed using ab200829, the same antibody clone in a different buffer formulation.

NFIB/NF1B2 + NFIC was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab200829 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab200829 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : HeLa whole cell lysate, 10 Âμg (Input).

Lane 2 : ab200829 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab200829 in HeLa whole cell lysate.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (<a href='/en-us/products/primary-antibodies/nfib-nf1b2-nfic-ctf-antibody-epr14504-ab200829'>ab200829</a>)

Observed band size: 45-65 kDa

false

• WB

Supplier Data

Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] - BSA and Azide free (AB251321)

This data was developed using ab200829, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Based on the sequence analysis, ab200829 recognizes six isoforms within human with predicted Mw’s of 56KDa, 55KDa, 48KDa, 45KDa, 48KDa and 49KDa, respectively.

All lanes:

Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (<a href='/en-us/products/primary-antibodies/nfib-nf1b2-nfic-ctf-antibody-epr14504-ab200829'>ab200829</a>) at 1/2000 dilution

Lane 1:

Mouse heart lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat brain lysate at 10 µg

Lane 5:

Rat heart lysate at 10 µg

Lane 6:

Rat spleen lysate at 10 µg

Lane 7:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 8:

RAW 264.7(Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 9:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 10:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 47 kDa,56 kDa

Observed band size: 45-65 kDa

false

Exposure time: 1min