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Product Info Summary
| SKU: | A04443-2 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human, Monkey, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-NF45/ILF2 Antibody Picoband™
SKU/Catalog Number
A04443-2
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-NF45/ILF2 Antibody Picoband™ catalog # A04443-2. Tested in ELISA, IF, IHC, ICC, WB, Flow Cytometry applications. This antibody reacts with Human, Monkey, 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-NF45/ILF2 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04443-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
IgG
Immunogen
E.coli-derived human Pyruvate Carboxylase/PC recombinant protein (Position: R88-D1165). Human PC shares 97% and 96.6% amino acid (aa) sequence identity with mouse and rat PC, respectively.
*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
A04443-2 is reactive to ILF2 in Human, Monkey, Mouse, Rat
Applications
A04443-2 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
43 kDa
Calculated molecular weight
52588 MW
Background of ILF2
Interleukin enhancer-binding factor 2 is a protein that in humans is encoded by the ILF2 gene. The protein encoded by this gene is a transcription factor required for T-cell expression of the interleukin 2 gene. It also binds RNA and is an essential component for encapsidation and protein priming of hepatitis B viral polymerase. The encoded 45 kDa protein (NF45, ILF2) forms a complex with the 90 kDa interleukin enhancer-binding factor 3 (NF90, ILF3), and this complex has been shown to affect the redistribution of nuclear mRNA to the cytoplasm, to repair DNA breaks by nonhomologous end joining, and to negatively regulate the microRNA processing pathway. Knockdown of NF45 or NF90 protein retards cell growth, possibly by inhibition of mRNA stabilization. Alternative splicing results in multiple transcript variants. Related pseudogenes have been found on chromosomes 3 and 14.
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.
Assay dilution & Images
Reconstitution
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, 0.1-0.25 µg/ml, Human, Monkey, Mouse, Rat
Immunohistochemistry, 1-2 µg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg /1x106 cells, Human
ELISA, 0.1-0.5 µg/ml, Human
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of NF45/ILF2 using anti-NF45/ILF2 antibody (A04443-2).
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 K562 whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: monkey COS-7 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat C6 whole cell lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse NIH/3T3 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 rabbit anti-NF45/ILF2 antigen affinity purified polyclonal antibody (Catalog # A04443-2) at 0.25 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for NF45/ILF2 at approximately 43 kDa. The expected band size for NF45/ILF2 is at 43 kDa.
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Figure 2. IHC analysis of NF45/ILF2 using anti-NF45/ILF2 antibody (A04443-2).
NF45/ILF2 was detected in a paraffin-embedded section of human esophageal squamous carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NF45/ILF2 Antibody (A04443-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Figure 3. IHC analysis of NF45/ILF2 using anti-NF45/ILF2 antibody (A04443-2).
NF45/ILF2 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NF45/ILF2 Antibody (A04443-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Figure 4. IHC analysis of NF45/ILF2 using anti-NF45/ILF2 antibody (A04443-2).
NF45/ILF2 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NF45/ILF2 Antibody (A04443-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Figure 5. IHC analysis of NF45/ILF2 using anti-NF45/ILF2 antibody (A04443-2).
NF45/ILF2 was detected in a paraffin-embedded section of human rectum adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NF45/ILF2 Antibody (A04443-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Figure 6. IHC analysis of NF45/ILF2 using anti-NF45/ILF2 antibody (A04443-2).
NF45/ILF2 was detected in a paraffin-embedded section of human thyroid papillary carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NF45/ILF2 Antibody (A04443-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 7. IHC analysis of NF45/ILF2 using anti-NF45/ILF2 antibody (A04443-2).
NF45/ILF2 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NF45/ILF2 Antibody (A04443-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 8. IHC analysis of NF45/ILF2 using anti-NF45/ILF2 antibody (A04443-2).
NF45/ILF2 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NF45/ILF2 Antibody (A04443-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Figure 9. IF analysis of NF45/ILF2 using anti-NF45/ILF2 antibody (A04443-2) and anti-Beta Tubulin antibody (M01857-3).
NF45/ILF2 was detected in immunocytochemical section of HELA cell. 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 rabbit anti-NF45/ILF2 Antibody (A04443-2) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 10. Flow Cytometry analysis of JK cells using anti-NF45/ILF2 antibody (A04443-2).
Overlay histogram showing JK cells stained with A04443-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NF45/ILF2 Antibody (A04443-2, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For ILF2 (Source: Uniprot.org, NCBI)
Gene Name
ILF2
Full Name
Interleukin enhancer-binding factor 2
Weight
52588 MW
Alternative Names
interleukin enhancer binding factor 2, 45kD; interleukin enhancer binding factor 2, 45kDa; interleukin enhancer-binding factor 2; MGC8391; NF45PRO3063; Nuclear factor of activated T-cells 45 kDa; nuclear factor of activated T-cells, 45-kDa ILF2 NF45, PRO3063 interleukin enhancer binding factor 2 interleukin enhancer-binding factor 2|interleukin enhancer binding factor 2, 45kD|interleukin enhancer binding factor 2, 45kDa|nuclear factor of activated T-cells, 45-kDa
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on ILF2, check out the ILF2 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for ILF2: 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].
Specific Publications For Anti-NF45/ILF2 Antibody Picoband™ (A04443-2)
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