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

Product Name

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

View all NFIB Antibodies

SKU/Catalog Number

M01537-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-NFIB/NF1B2 Antibody Picoband™ (monoclonal, 4D6E4) catalog # M01537-1. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

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.

Cite This Product

Anti-NFIB/NF1B2 Antibody Picoband™ (monoclonal, 4D6E4) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01537-1)

Host

Mouse

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

Clonality

Monoclonal

Clone Number

4D6E4

Isotype

Mouse IgG2b

Immunogen

A synthetic peptide corresponding to a sequence in the middle region of human NFIB/NF1B2, identical to the related mouse and rat sequences.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

M01537-1 is reactive to NFIB in Human, Mouse, Rat

Applications

M01537-1 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

68 kDa

Calculated molecular weight

47.442kDa

Background of NFIB

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.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human, Mouse, Rat

Validation Images & Assay Conditions

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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.

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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.

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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.

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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.

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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.

Gene/Protein Information For NFIB (Source: Uniprot.org, NCBI)

Alternative Names

CCAAT-box-binding transcription factor; CTF; HMGIC/NFIB; NF-I/B; NFI-B; NFIB2; NFIB3; NFI-RED; nuclear factor 1 B-type; Nuclear factor 1/B; nuclear factor I/BNF1-B; TGGCA-binding protein NFIB CTF, HMGIC/NFIB, MACID, NF-I/B, NF1-B, NFI-B, NFI-RED2, NFIB3, NFIB nuclear factor I B nuclear factor 1 B-type|CCAAT-box-binding transcription factor|TGGCA-binding protein|nuclear factor 1/B

For more info on NFIB, check out the NFIB Infographic

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In this infographic, you will see the following information for NFIB: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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