Antibodies, Proteins, Kits and Reagents for Life Science

Anti-NFIB / NF1B2+NFIC antibody [EPR14504] - BSA and Azide free (ab251321)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR14504] to NFIB / NF1B2+NFIC - BSA and Azide free
Suitable for: WB, Flow Cyt, IP, IHC-P, ICC/IF
Reacts with: Mouse, Rat, Human

Product name

Anti-NFIB / NF1B2+NFIC antibody [EPR14504] - BSA and Azide free
See all NFIB / NF1B2+NFIC primary antibodies
Description

Rabbit monoclonal [EPR14504] to NFIB / NF1B2+NFIC - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

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

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
General notes

Ab251321 is the carrier-free version of ab200829. 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
ab251321 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C. Do Not Freeze.
Storage buffer

pH: 7.2
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Clonality

Monoclonal
Clone number

EPR14504
Isotype

IgG
Research areas

Western blot - Anti-NFIB / NF1B2+NFIC antibody [EPR14504] - BSA and Azide free (ab251321)

All lanes : Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829) at 1/10000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 56, 47 kDa
Observed band size: 45-65 kDa
why is the actual band size different from the predicted?

Exposure time: 30 seconds

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.
Immunocytochemistry - Anti-NFIB / NF1B2+NFIC 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2+NFIC 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 Cytometry - Anti-NFIB / NF1B2+NFIC antibody [EPR14504] - BSA and Azide free (ab251321)

This data was developed using ab200829, the same antibody clone in a different buffer formulation.
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.
Immunoprecipitation - Anti-NFIB / NF1B2+NFIC 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2+NFIC 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2+NFIC 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.
Western blot - Anti-NFIB / NF1B2+NFIC antibody [EPR14504] - BSA and Azide free (ab251321)

Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829) at 1/10000 dilution + HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 20 µg

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

Predicted band size: 56, 47 kDa
Observed band size: 45-65 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

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.
Western blot - Anti-NFIB / NF1B2+NFIC antibody [EPR14504] - BSA and Azide free (ab251321)

Lanes 1-8 : Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829) at 1/2000 dilution
Lanes 9-10 : Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829) at 1/1000 dilution

Lane 1 : Mouse heart lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat heart lysate
Lane 6 : Rat spleen lysate
Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
Lane 8 : RAW 264.7(Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 56, 47 kDa
Observed band size: 45-65 kDa why is the actual band size different from the predicted?

Exposure time: 1 minute

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.
Anti-NFIB / NF1B2+NFIC antibody [EPR14504] - BSA and Azide free (ab251321)

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