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Anti-NFIB/NF1B2 Antibody Picoband™ (monoclonal, 4D6E4)

Figure 1. Western blot analysis of NFIB/NF1B2 using anti-NFIB/NF1B2 antibody (M01537-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human 293T whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-NFIB/NF1B2 antigen affinity purified monoclonal antibody (Catalog # M01537-1) at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for NFIB/NF1B2 at approximately 68 kDa. The expected band size for NFIB/NF1B2 is at 68 kDa.
Figure 2. IF analysis of NFIB/NF1B2 using anti-NFIB/NF1B2 antibody (M01537-1).
NFIB/NF1B2 was detected in an immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL mouse anti-NFIB/NF1B2 Antibody (M01537-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 3. Flow Cytometry analysis of A431 cells using anti-NFIB/NF1B2 antibody (M01537-1).
Overlay histogram showing A431 cells stained with M01537-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-NFIB/NF1B2 Antibody (M01537-1, 1 µg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 4. Flow Cytometry analysis of C6 cells using anti-NFIB/NF1B2 antibody (M01537-1).
Overlay histogram showing C6 cells stained with M01537-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-NFIB/NF1B2 Antibody (M01537-1, 1 µg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 5. Flow Cytometry analysis of Neuro-2a cells using anti-NFIB/NF1B2 antibody (M01537-1).
Overlay histogram showing Neuro-2a cells stained with M01537-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-NFIB/NF1B2 Antibody (M01537-1, 1 µg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IF, ICC, FC
Primary Accession	O00712
Host	Mouse
Isotype	Mouse IgG2b
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Lyophilized
Description	Anti-NFIB/NF1B2 Antibody Picoband™ (monoclonal, 4D6E4) . Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Reconstitution	Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Additional Information
Gene ID	4781
Other Names	Nuclear factor 1 B-type, NF1-B, Nuclear factor 1/B, CCAAT-box-binding transcription factor, CTF, Nuclear factor I/B, NF-I/B, NFI-B, TGGCA-binding protein, NFIB
Calculated MW	47442 Da
Application Details	Western blot, 0.25-0.5 µg/ml, Human Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human Flow Cytometry, 1-3 µg/1x10^6 cells, Human, Mouse, Rat
Source	Eukaryota
Contents	Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clone Names	Clone: 4D6E4
Immunogen	A synthetic peptide corresponding to a sequence in the middle region of human NFIB/NF1B2, identical to the related mouse and rat sequences.
Purification	Immunogen affinity purified.
Storage	At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	NFIB
Function	Transcriptional activator of GFAP, essential for proper brain development (PubMed:30388402). Recognizes and binds the palindromic sequence 5'-TTGGCNNNNNGCCAA-3' present in viral and cellular promoters and in the origin of replication of adenovirus type 2. These proteins are individually capable of activating transcription and replication.
Cellular Location	Nucleus.

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

Nuclear factor 1 B-type is a protein that in humans is encoded by the NFIB gene. The NFIB gene is a part of the NFI gene complex that includes three other genes (NFIA, NFIC and NFIX). The NFIB gene is a protein coding gene that also serves as a transcription factor. This gene is essential in embryonic development and it works together with its gene complex to initiate tissue differentiation in the fetus. Through knockout experiments, researchers found that mice without the NFIB gene have severely underdeveloped lungs. This mutation does not seem to cause spontaneous abortions because in utero the fetus does not use its lungs for respiration. However, this becomes lethal once the fetus is born and has to take its first breath. It is thought that NFIB plays a role in down regulating the transcription factors TGF-β1 and Shh in normal gestation because they remained high in knockout experiments. The absence of NFIB also leads to insufficient amounts of surfactant being produced which is one reason why the mice cannot breathe once it is born. The knockout experiments demonstrated that NFIB has a significant role in fore-brain development. NFIB is typically found in pontine nuclei of the CNS, the cerebral cortex and the white matter of the brain and without NFIB these areas are dramatically affected.

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¥ 3,367.00

Cat# ABO15119

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100 µg

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Anti-NFIB/NF1B2 Antibody Picoband™ (monoclonal, 4D6E4)

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