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Rat Heterogeneous nuclear ribonucleoproteins A2/B1 (Hnrnpa2b1) ELISA Kit (RTEB0567)

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SKU:
RTEB0567
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
A7VJC2
ELISA Type:
Sandwich
Reactivity:
Rat
$779

Description

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Rat Heterogeneous nuclear ribonucleoproteins A2/B1 (Hnrnpa2b1) ELISA Kit

The Rat Heterogeneous Nuclear Ribonucleoproteins A2/B1 (HNRNPA2B1) ELISA Kit is a valuable tool for researchers looking to accurately measure levels of HNRNPA2B1 in rat samples. This kit offers high sensitivity and specificity, ensuring precise and reproducible results in serum, plasma, and cell culture supernatants.HNRNPA2B1 is a key protein involved in RNA processing and regulation, playing a crucial role in gene expression and cellular function.

Dysregulation of HNRNPA2B1 has been linked to various diseases, including cancer and neurodegenerative disorders, making it a valuable biomarker for researchers studying these conditions and exploring potential therapeutic interventions.Overall, the Rat HNRNPA2B1 ELISA Kit is a powerful tool for investigating the role of HNRNPA2B1 in disease pathogenesis and for developing new strategies for diagnosis and treatment.

Product Name:Rat Heterogeneous nuclear ribonucleoproteins A2/B1 (Hnrnpa2b1) ELISA Kit
SKU:RTEB0567
Size:96T
Target:Rat Heterogeneous nuclear ribonucleoproteins A2/B1 (Hnrnpa2b1)
Synonyms:hnRNP A2/B1, Hnrnp, Hnrpa2b1
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Rat
Detection Range:0.312-20ng/mL
Sensitivity:0.16ng/mL
Intra CV:Provided with the Kit
Inter CV:Provided with the Kit
Linearity:Provided with the Kit
Recovery:Provided with the Kit
Function:Heterogeneous nuclear ribonucleoprotein (hnRNP) that associates with nascent pre-mRNAs, packaging them into hnRNP particles. The hnRNP particle arrangement on nascent hnRNA is non-random and sequence-dependent and serves to condense and stabilize the transcripts and minimize tangling and knotting. Packaging plays a role in various processes such as transcription, pre-mRNA processing, RNA nuclear export, subcellular location, mRNA translation and stability of mature mRNAs. Forms hnRNP particles with at least 20 other different hnRNP and heterogeneous nuclear RNA in the nucleus (PubMed:19099192). Involved in transport of specific mRNAs to the cytoplasm in oligodendrocytes and neurons: acts by specifically recognizing and binding the A2RE (21 nucleotide hnRNP A2 response element) or the A2RE11 (derivative 11 nucleotide oligonucleotide) sequence motifs present on some mRNAs, and promotes their transport to the cytoplasm (PubMed:9578590, PubMed:10567417). Specifically binds single-stranded telomeric DNA sequences, protecting telomeric DNA repeat against endonuclease digestion (PubMed:15659580). Also binds other RNA molecules, such as primary miRNA (pri-miRNAs): acts as a nuclear 'reader' of the N6-methyladenosine (m6A) mark by specifically recognizing and binding a subset of nuclear m6A-containing pri-miRNAs. Binding to m6A-containing pri-miRNAs promotes pri-miRNA processing by enhancing binding of DGCR8 to pri-miRNA transcripts. Involved in miRNA sorting into exosomes following sumoylation, possibly by binding (m6A)-containing pre-miRNAs. Acts as a regulator of efficiency of mRNA splicing, possibly by binding to m6A-containing pre-mRNAs.
Uniprot:A7VJC2
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant rat Heterogeneous nuclear ribonucleoproteins A2/B1
Sub Unit:Identified in the spliceosome C complex. Identified in a IGF2BP1-dependent mRNP granule complex containing untranslated mRNAs. Interacts with IGF2BP1. Interacts with C9orf72. Interacts with DGCR8. Interacts with TARDBP. Interacts with CKAP5.
Research Area:Epigenetics
Subcellular Location:Nucleus Nucleoplasm Cytoplasm Cytoplasmic granule Secreted Exosome Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Component of ribonucleosomes. Not found in the nucleolus. Found in exosomes follwong sumoylation.
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:hnRNP A2/B1: Involved with pre-mRNA processing. Forms complexes (ribonucleosomes) with at least 20 other different hnRNP and heterogeneous nuclear RNA in the nucleus. Identified in the spliceosome C complex. Identified in a mRNP granule complex, at least composed of ACTB, ACTN4, DHX9, ERG, HNRNPA1, HNRNPA2B1, HNRNPAB, HNRNPD, HNRNPL, HNRNPR, HNRNPU, HSPA1, HSPA8, IGF2BP1, ILF2, ILF3, NCBP1, NCL, PABPC1, PABPC4, PABPN1, RPLP0, RPS3, RPS3A, RPS4X, RPS8, RPS9, SYNCRIP, TROVE2, YBX1 and untranslated mRNAs. Interacts with IGF2BP1. 2 isoforms of the human protein are produced by alternative splicing.Protein type: RNA-binding; RNA splicing; SpliceosomeCellular Component: nucleoplasm; spliceosome; neuron projection; membrane; cell soma; cytoplasm; perikaryon; chromatin; nucleus; ribonucleoprotein complexMolecular Function: nucleic acid binding; single-stranded telomeric DNA binding; RNA binding; nucleotide bindingBiological Process: negative regulation of nuclear mRNA splicing, via spliceosome; RNA splicing; male gonad development; oligodendrocyte differentiation; negative regulation of transcription from RNA polymerase II promoter; response to mineralocorticoid stimulus; mRNA processing; lung development; RNA transport
UniProt Protein Details:
NCBI Summary:
UniProt Code:A7VJC2
NCBI GenInfo Identifier:157265559
NCBI Gene ID:362361
NCBI Accession:NP_001098083.1
UniProt Secondary Accession:A7VJC2,A7VJC1, A7VJC3, A7VJC4
UniProt Related Accession:A7VJC2
Molecular Weight:33,959 Da
NCBI Full Name:heterogeneous nuclear ribonucleoproteins A2/B1
NCBI Synonym Full Names:heterogeneous nuclear ribonucleoprotein A2/B1
NCBI Official Symbol:Hnrnpa2b1
NCBI Official Synonym Symbols:hnRNP; Hnrpa2; Hnrpa2b1
NCBI Protein Information:heterogeneous nuclear ribonucleoproteins A2/B1
UniProt Protein Name:Heterogeneous nuclear ribonucleoproteins A2/B1
UniProt Synonym Protein Names:
Protein Family:Heterogeneous nuclear ribonucleoproteins
UniProt Gene Name:Hnrnpa2b1
UniProt Entry Name:ROA2_RAT
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir

*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatant Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysates Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenates The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysates Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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