Anti-NFIB / NF1B2 antibody [EPR14122]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2 antibody [EPR14122] (AB186738)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling NFIB / NF1B2 with ab186738 at a 1/500 dilution. Counter stained with hematoxylin.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NFIB / NF1B2 antibody [EPR14122] (AB186738)

Immunofluorescence analysis of paraformaldehyde-fixed HeLa cells labeling NFIB / NF1B2 with ab186738 at a 1/100 dilution. As secondary antibody goat anti-rabbit IgG (Alexa Fluor®555) was used at a 1/200. In blue DAPI staining.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NFIB / NF1B2 antibody [EPR14122] (AB186738)

ab186738 staining NFIB / NF1B2 in the human cell line HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permiabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/70. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control : Rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

• IP

Unknown

Immunoprecipitation - Anti-NFIB / NF1B2 antibody [EPR14122] (AB186738)

Western blot analysis on immunoprecipitation from 1) Human fetal brain lysate and 2) PBS, labeling NFIB / NF1B2 using ab186738 at 1/50 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG at a 1/1500 dilution.

All lanes:

Immunoprecipitation - Anti-NFIB / NF1B2 antibody [EPR14122] (ab186738)

Predicted band size: 47 kDa

Observed band size: 47 kDa

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• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2 antibody [EPR14122] (AB186738)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling NFIB / NF1B2 with ab186738 at a 1/500 dilution. Counter stained with hematoxylin.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-NFIB / NF1B2 antibody [EPR14122] (AB186738)

All lanes:

Western blot - Anti-NFIB / NF1B2 antibody [EPR14122] (ab186738) at 1/5000 dilution

All lanes:

HeLa 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

Observed band size: 47 kDa,53 kDa,55 kDa

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• WB

Lab

Western blot - Anti-NFIB / NF1B2 antibody [EPR14122] (AB186738)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of HeLa whole cell lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab186738 (1/5000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/20,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique. ab186738 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant for temperatures ranging between 4°C, 22°C and 37°C, demonstrating that antibody activity is not affected under these conditions.

All lanes:

Western blot - Anti-NFIB / NF1B2 antibody [EPR14122] (ab186738) at 1/5000 dilution

All lanes:

HeLa whole cell lysate at 15 µg with NDFM/TBST

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 1/20000 dilution

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Exposure time: 20s

• WB

Supplier Data

Western blot - Anti-NFIB / NF1B2 antibody [EPR14122] (AB186738)

All lanes:

Western blot - Anti-NFIB / NF1B2 antibody [EPR14122] (ab186738) at 1/5000 dilution

All lanes:

Human fetal brain 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-ab136636'>ab136636</a>) at 1/1000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

true

• WB

Supplier Data

Western blot - Anti-NFIB / NF1B2 antibody [EPR14122] (AB186738)

All lanes:

Western blot - Anti-NFIB / NF1B2 antibody [EPR14122] (ab186738) at 1/1000 dilution

Lane 1:

C6 cell lysate at 10 µg

Lane 2:

PC-12 cell lysate at 10 µg

Lane 3:

NIH/3T3 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

Observed band size: 47 kDa

true