hnRNP A1 Antibody (4B10) - BSA Free

Western Blot: hnRNP A1 Antibody (4B10) [NB100-672] - Western blot shows lysates of HEK293T human embryonic kidney parental cell line and hnRNP A1 knockout (KO) HEK293T cell line. PVDF membrane was probed with 0.5 ug/ml of Mouse Anti-Human hnRNP A1 Monoclonal Antibody (Catalog # NB100-672) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog #HAF018). Specific band was detected for hnRNP A1 at approximately 35 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in the knockout HEK293T cell line. This experiment was conducted under reducing conditions.

Western Blot: hnRNP A1 Antibody (4B10) [NB100-672] - Whole cell protein from HeLa (lane 1), A431 (lane 2), Hek293 (lane 3) and MCF7 (lane 4) were separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 0.5 mg/ml anti-hnRNP in 1% milk. Precision Plus Protein All Blue molecular weight markers (BioRad) were detected with 1 ug/ml Anti-Blue Marker antibody (NBP2-33376). Both antibodies were detected with an anti-mouse HRP secondary antibody using chemiluminescence.

Immunocytochemistry/Immunofluorescence: hnRNP A1 Antibody (4B10) [NB100-672] - The hnRNP A1 (4B10) antibody was tested in HeLa cells at a 1:200 dilution against Dylight 488 (Green). Actin and nuclei were counterstained against Phalloidin 568 (Red) and DAPI (Blue), respectively.

Western Blot: hnRNP A1 Antibody (4B10) [NB100-672] - Cell line lysates were separated on SDS-PAGE and probed with 0.5 ug/mL Monoclonal Anti-hnRNP-A1 Clone: 4B10 (NB100-672). Lanes: 1-HeLa, 2-Jurkat, 3-HEK-293T, 4-WiDr, 5-NCI-H1299, 6-F9.

Western blot shows lysates of HEK 293T cell line and hnRNP A1 knockout HEK 293T cell line (KO). Nitrocellulose membrane was probed with hnRNP A1 Antibody (Catalog # NB100-672) O/N at 4C, followed by HRP-conjugated Secondary Antibody and ECL detection. A specific band was detected for hnRNP A1 (as indicated) in the parental HAP1 cell line, but is not detectable in knockout HAP1 cell line. The Ponceau stained transfer of the blot is shown. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

HEK 293T WT and hnRNP A1 KO cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with hnRNP A1 antibody and with the corresponding Alexa-fluor 555 coupled secondary antibody including DAPI. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative images of the blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Antibody dilution used: 1/1000. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

• Reactivity: Hu, Mu, Bv, Ca
• Applications: WB, ELISA, ICC/IF, IHC, IP, MA, KD
• Clone: 4B10
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

hnRNP A1 Antibody (4B10) - BSA Free Summary

• Immunogen: Full length partially purified human hnRNP A1. [UniProt# P09651]
• Localization: Nucleus. Cytoplasm. Shuttles continuously between the nucleus and the cytoplasm along with mRNA. Component of ribonucleosomes.
• Specificity: Does not react with hnRNP A2 or B
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: HNRNPA1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • ELISA Indirect
  • Immunocytochemistry/ Immunofluorescence 5 ug/ml
  • Immunohistochemistry reported in scientific literature (PMID 3129338)
  • Immunoprecipitation 1:10-1:500
  • Knockdown Validated
  • Microarray
  • Western Blot 0.25 - 0.5 ug/ml
• Application Notes: In WB a band can be seen between 32-35 kDa. This antibody immunoprecipitates the hnRNP complex.
• Control:

  A-431 Whole Cell Lysate
  HeLa Whole Cell Lysate
  HEK293 Cell Lysate
  MCF-7 Cell Lysate

Reactivity Notes

Not yet tested in other species.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for hnRNP A1 Antibody (4B10) - BSA Free

• Helix-destabilizing protein
• heterogeneous nuclear ribonucleoprotein A1
• heterogeneous nuclear ribonucleoprotein A1B protein
• heterogeneous nuclear ribonucleoprotein B2 protein
• heterogeneous nuclear ribonucleoprotein core protein A1
• hnRNP A1
• hnRNP core protein A1
• hnRNPA1
• hnRNP-A1
• HNRPA1MGC102835
• nuclear ribonucleoprotein particle A1 protein
• single-strand DNA-binding protein UP1
• Single-strand RNA-binding protein

Background

RNA polymerase II trans cripts in the nucleus are in complex with several proteins called heterogeneous nuclear ribonucleoproteins (hnRNPs). These proteins are important in biological activities such as transcription, premRNA processing, cytoplasmic mRNA translation, and turnover. hnRNPs can be isolated either by immunoprecipitation or by sucrose gradient fractionation of cell extracts.2, 4, 5 Isolated hnRNPs consist of protein groups named A to U and many of these protein groups consist of more than one isoform. The major steady-state proteins of the isolated hnRNP complex are A1, A2, B1, B2, C1, and C2, with a range of molecular weight starting with 34 kDa up to 43 kDa.2, 4, 5 hnRNP-A1 is important in pre-mRNA processing and in mRNA export from the nucleus. The protein binds to its RNA target through a consensus RNA-binding site UAGGGU. The protein contains a 38-amino acid domain called M9, which is important for the interaction with the transportin protein and therefore, for its import and export from the nucleus. RanGTP mediates dissociation of hnRNP -A1 from transportin. hnRNP-A1 is ubiquitously expressed with a higher expression in proliferating and/or transformed cells than in differentiated tissues.6

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for hnRNP A1 Antibody (NB100-672)(24)

Publications using NB100-672

• Huelga SC, Vu AQ, Arnold JD et al. Integrative genome-wide analysis reveals cooperative regulation of alternative splicing by hnRNP proteins Cell Rep 2012-02-01 [PMID: 22574288] (Human)
• Bonnal S, Pileur F, Orsini C et al. Heterogeneous nuclear ribonucleoprotein A1 is a novel internal ribosome entry site trans-acting factor that modulates alternative initiation of translation of the fibroblast growth factor 2 mRNA. J Biol Chem. 2005-02-11 [PMID: 15525641] (WB, IP, Human)
• Pino I, Pio R, Toledo G et al. Altered patterns of expression of members of the heterogeneous nuclear ribonucleoprotein (hnRNP) family in lung cancer. Lung Cancer. 2003-08-01 [PMID: 12871776] (WB, ICC/IF, Human)
• Guil S, Gattoni R, Carrascal M et al. Roles of hnRNP A1, SR proteins, and p68 helicase in c-H-ras alternative splicing regulation. Mol Cell Biol. 2003-04-01 [PMID: 12665590] (WB, Human)
• Mili S, Shu HJ, Zhao Y et al. Distinct RNP complexes of shuttling hnRNP proteins with pre-mRNA and mRNA: candidate intermediates in formation and export of mRNA. Mol Cell Biol. 2001-11-01 [PMID: 11585913] (WB, Human)
• van der Houven van Oordt W, Diaz-Meco MT, Lozano J et al. The MKK(3/6)-p38-signaling cascade alters the subcellular distribution of hnRNP A1 and modulates alternative splicing regulation. J Cell Biol. 2000-04-17 [PMID: 10769024] (ICC/IF, IP, WB, Mouse)
• Del Gatto-Konczak F, Olive M, Gesnel MC, Breathnach R. hnRNP A1 recruited to an exon in vivo can function as an exon splicing silencer. Mol Cell Biol. 1999-01-01 [PMID: 9858549] (IP, Human)
• Siomi MC, Eder PS, Kataoka N et al. Transportin-mediated nuclear import of heterogeneous nuclear RNP proteins. J Cell Biol. 1997-09-22 [PMID: 9298975] (WB, ICC/IF, IP, Human)
• Pinol-Roma S, Choi YD, Matunis MJ, Dreyfuss G. Immunopurification of heterogeneous nuclear ribonucleoprotein particles reveals an assortment of RNA-binding proteins. Genes Dev. 1988-02-01 [PMID: 3129338] (IP, WB, ELISA, ICC/IF, Human)
• Pinol-Roma S, Swanson MS, Gall JG, Dreyfuss G. A novel heterogeneous nuclear RNP protein with a unique distribution on nascent transcripts. J Cell Biol. 1989-12-01 [PMID: 2687284] (ICC/IF, Human)
• Pinol-Roma S, Dreyfuss G. Transcription-dependent and transcription-independent nuclear transport of hnRNP proteins. Science. 1991-07-19 [PMID: 1857966] (ICC/IF, Human)
• Halbert D, Domenyuk V, Spetzler D et al. Aptamers and uses thereof United States Patent Application US 9958448 B2 2018-01-01
• Chen Y, Wu Y, Li J et al. Cooperative regulation of Zhx1 and hnRNPA1 drives the cardiac progenitor-specific transcriptional activation during cardiomyocyte differentiation Cell death discovery 2023-07-14 [PMID: 37452012] (WB, Mouse)
• Yasuhara T, Xing Y, Bauer N et al. Condensates induced by transcription inhibition localize active chromatin to nucleoli Molecular Cell 2022-06-01 [PMID: 35662392] (Immunocytochemistry/ Immunofluorescence, Human)
• Fang MY, Markmiller S, Vu AQ, et al. Small-Molecule Modulation of TDP-43 Recruitment to Stress Granules Prevents Persistent TDP-43 Accumulation in ALS/FTD Neuron 2019-06-18 [PMID: 31272829]
• Marrone, L;Drexler, HCA;Wang, J;Tripathi, P;Distler, T;Heisterkamp, P;Anderson, EN;Kour, S;Moraiti, A;Maharana, S;Bhatnagar, R;Belgard, TG;Tripathy, V;Kalmbach, N;Hosseinzadeh, Z;Crippa, V;Abo-Rady, M;Wegner, F;Poletti, A;Troost, D;Aronica, E;Busskamp, V; FUS pathology in ALS is linked to alterations in multiple ALS-associated proteins and rescued by drugs stimulating autophagy Acta Neuropathol. 2019-04-01 [PMID: 30937520] (ICC/IF, Human)
• Thompson VF, Victor RA, Morera AA et al. Transcription-Dependent Formation of Nuclear Granules Containing FUS and RNA Pol II Biochemistry 2018-12-26 [PMID: 30488693] (Immunocytochemistry/ Immunofluorescence, Human)
• Gasperini L, Rossi A, Cornella N et al. The hnRNP raly regulates PRMT1 expression and interacts with the ALS-linked protein FUS: implication for reciprocal cellular localization. Mol. Biol. Cell. 2018-10-24 [PMID: 30354839] (WB, Human)
• Lampe S, Kunze M, Scholz A et al. Identification of the TXNIP IRES and characterization of the impact of regulatory IRES trans-acting factors. Biochim Biophys Acta Gene Regul Mech 2018-02-01 [PMID: 29378331] (WB)
• Hu J, Khodadadi-Jamayran A, Mao M et al. AKAP95 regulates splicing through scaffolding RNAs and RNA processing factors. Nat Commun. 2016-11-08 [PMID: 27824034]
• Kamelgarn M, Chen J, Kuang L et al. Proteomic analysis of FUS interacting proteins provides insights into FUS function and its role in ALS Biochim. Biophys. Acta 2016-07-25 [PMID: 27460707] (WB)
• Kunze MM, Benz F, Braub TF et al. sST2 translation is regulated by FGF2 via an hnRNP A1-mediated IRES-dependent mechanism. Biochim. Biophys. Acta. 2016-05-08 [PMID: 27168114] (ICC/IF, WB, IP, Human)
• Fujiya M, Konishi H, Mohamed Kamel MK et al. microRNA-18a induces apoptosis in colon cancer cells via the autophagolysosomal degradation of oncogenic heterogeneous nuclear ribonucleoprotein A1 Oncogene 2013-10-28 [PMID: 24166503] (WB, Human)
  
  Details:
  This citation used the Alkaline Phosphatase version of this antibody.
• Lin, Y-T, Yen, P H. A novel nucleocytoplasmic shuttling sequence of DAZAP1, a testis-abundant RNA-binding protein, . 12, 1486-1493. 2006-01-01 [PMID: 16772659]

Bioinformatics

• Gene Symbol: HNRNPA1
• Entrez:
  • 3178(Human)
• Uniprot:
  • P09651(Human)