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We validate the specificity of these antibodies to EIF4A2 by testing them on tissues known to express EIF4A2 positively and negatively. Browse below to find the EIF4A2 antibody that suites your experiment. We have 3 of these antibodies and many publications and validation images.
If you cannot find antibodies that fit your needs, contact us for making custom antibodies. We have a full suite of custom antibody services covering from research to diagnostic and therapeutic applications.
Facts about Eukaryotic initiation factor 4A-II.
|DEAD box helicase family|
ATP-dependent RNA helicase eIF4A-2; BM-010; DDX2Beukaryotic translation initiation factor 4A, isoform 2; EC 3.6.1; EC 188.8.131.52; EIF4A; eIF-4A-II; eIF4A-II; EIF4F; eukaryotic initiation factor 4A-II; eukaryotic translation initiation factor 4A; eukaryotic translation initiation factor 4A2
|Sequence:||3; NC_000003.12 (186783577..186789897)|
Boster Bio may have the antibody you need to determine the most effective uses for the EIF4A2 marker. This antibody reacts to Human, Mouse, and Rat cells. You might also be interested in understanding how to use EIF4A2 marker for your experiments. This article will go over some of the most effective applications for this marker.
The mechanism of the TGEV replication cycle was revealed through the discovery of the EIF4A2 marker. Human EIF4A2 is a receptor for RdRp NS5B from the hepatitis Cvirus to assist in genome RNA synthesis process for the NS5B protein. Therefore, EIF4A2 can be considered a new therapeutic target that could help TGEV replication.
TGEV is a highly contagious disease causing acute diarrhea in piglets. This is a significant economic problem and identifying host proteins that interact with viral proteins is essential to understanding the pathogenesis the virus. Seven protein molecules from the cell have been identified as potential binding partners for the TGEVM protein by researchers. These proteins are involved in many processes such as transcription of genes, folding proteins and metabolism. In the intestinal cells of piglets, EIF4A2, a novel M ligand, was identified.
To alter the EIF4A2 gene, siRNA2 was transfected into pIECs. TGEV infection was carried out for 48 hours in pIECs. After infection, total RNA was isolated from the cells by using RNAiso plus and reverse transcription. The b-actin gene served as an internal standard. The resultant RNA samples were then subjected to quantitative PCR using a two-cycle threshold method.
Co-IP assays are performed using IP kits KIP-1 (Protientech, China) and pIECs infected with TGEV or control cells were harvested 24 hours after infection. After each wash cells were sprayed with paraformaldehyde (4%) and allowed to dry at the room temperature for 15 mins. Afterward, pIECs were blotted using 5% bovine serum albumin. Then, a specific pAb against EIF4A2 was added to lysate supernate overnight at 4°C.
The EIF4A2 proteins performs two purposes. One is a binding partner for miRNAs and the second is an activator for suppression via RNA. While both functions are essential for the proper functioning of the virus, they could have distinct roles. Certain viruses may also undergo mutations to block the inhibition of cellular microRNAs. However, future studies will confirm this association and determine the precise role EIF4A2 plays in the process of repression by viruses.
To determine the role played by EIF4A2 in the repression of transcription in viral cells we performed a co-IP assay. We transfected pIECs using siRNA2 against EIF4A2 and inoculated them TGEV for 48 hours. The total RNA was extracted and reverse-transcribed using PrimerScript(tm) Master RT Mix. For internal control the b-actin gene is utilized. The resulting IP Lysate has a total of one to two mg of total protein. The Reed and Muench method was used to calculate the TCID50.
RNA binding affinity of eIF4A1 and eIF4A2 is different. eIF4A1 has a low affinity for dsRNA, whereas eIF4A2 has a strong preference for purine-only RNA. Both eIF4A1 and eIF4A2 have a similar affinity in RNA binding.
These antibodies have been verified by using an ELISA to identify viral antibodies in the immune system. Numerous researchers have used our boster antibody, which are highly specific over the past 25-years. In addition, our antibodies have been validated on Western Blotting, immunohistochemistry, and ELISA. We are very proud of our antibodies as well as the quality of their testing.
For the detection of protein expression, the quantitative PCR (qPCR), method is extensively employed. While the EIF4A2 gene interacts with CNOT1 however, the interaction is not carried out by RNA. The N-terminus of the gene is susceptible to major sequence divergence. To differentiate between high and low levels of expression, a cutoff value was set at 3.5.
To determine if eIF4A2 could be an important co-factor in initiation of translation, we employed the global translation initiation sequence (GRIP-Seq) in HEK293 cells. Lactimidomycin-treated cells were used to obtain peaks from initiating ribosomes. eIF4A2-bound MRNAs are likely to possess active uORFs. A subset, if any, of mRNAs are translated from an annotated AUG start codon Other mRNAs are able to initiate ribosomes located at downstream and upstream start sites.
The amplification of the cDNA fragment was completed with qPCR. The target gene (EIF4A2) is a protein that regulates a variety of cell functions and is produced by many cells of the body. The protein levels in the samples can't be determined until they have been reduced tenfold. This is a reliable method to determine the amount of protein in the sample.
QPCR is a combination of two PCR primers and the probe. The probe anneals into the middle of the amplicon and is accompanied by an molecule called a reporter. The reporter molecule is then identified by a quencher molecule that blocks fluorescence. The results of the amplification process are plotted against the threshold cycle number. These results can then be used to calculate the amount of the molecule targeted for amplification.
eIF4A activity has been linked to cancer, as well as its role as a transcription initiator. In addition, eIF4A1 expression correlates with cell proliferation, however, eIF4A2 expression is not. The analysis of GO-term enrichment also revealed significant functional differences between these eIF4A paralogs. Both proteins bind mRNAs that encode signaling factors. However, eIF4A1-bound mRNAs weren't enhanced for any particular term.
Researchers have identified a novel M protein ligand, EIF4A2, in the intestinal cells of pigs. The knockdown of EIF4A2 lowered M protein proliferativeness and decreased TGEV replication in DF1 cells. These findings could provide insights into the coronavirus replication process, and could be useful in the development of new strategies for treating coronavirus. The novel EIF4A2 ligand for M protein was also discovered by the researchers in intestinal cells. This is a discovery that had not been made in previous studies.
pIECs were transfected with siRNA2 against EIF4A2. Inoculated with TGEV for 48 h PIECs were then lysed with RNAiso plus. Then, the total RNA was extracted. Quantitative PCR was done using PrimerScript(tm) RT Master Mix with the b-actin gene used as an internal control. Two-Ct was employed to measure the expression of the gene in relation to.
To test this, pIECs infected by TGEV were allowed to live for up to 48 hours before being washed three times using PBS. The pIECs were infected with TGEV for 15 minutes at the room temperature. After that, they were fixed in paraformaldehyde (4 percent) and permeated with 0.3% Triton X-100 (10 min). After that, the supernatant was removed and blotted with bovine serum albumin, 5. IP lysis buffer was then added to the pIECs overnight at 4 degC. The IP Lysate was then supernatated to rabbit pAb against EIF4A2 then was incubated with 1:100 TGEV protein for further detection.
Boster researchers may submit their results for species and applications using Boster's ELISA kit. If they have positive results, they may be qualified for product credits. These credits are open to all scientists, whether they are located in the United States, China, or any other country. If you're an academic, you can submit your research results for review, and you could be awarded product credits for using Boster's kit.
Boster Bio has over 16,000 different antibodies in its catalog All of which have been thoroughly validated and tested. These antibodies can be used in multiple applications like IHC, WB and western Blotting. Boster also provides kits and rabbit polyclonal antibodies. Boster also offers a range of reagents such as buffers and lysates aswell as solutions and controls.
Dual labeling is a great way to improve analysis and research. The results of both antibodies targeting the same protein are stronger which allows scientists to explore more questions with a single specimen. Furthermore, it allows researchers to gain more context and better be aware of the interrelationship between different markers. Boster's primary antibodies with high affinity have this benefit. But how do we know which are the best?
First, the right amount of antibody is required to achieve a specific immunocytochemistry result. The ideal antibody must be able to bind to a specific antigen. To avoid binding that is not specific, it must be dilute to a high concentration. Finally, it must be accessible to the specific antibody used. A lack of adequate documentation on antibody controls could result in errors that can be repeated.
Dual antibody detection is more sensitive, however it requires more work to avoid cross-reactivity. This is due to the fact that the primary antibody contains epitopes that are shared with secondary antibodies. Dual antibodies can be used together to detect the target protein. Dual-antibody detection is more complicated and requires optimization. Additionally, dual-antibody detection is sensitive, but it requires more labeling.