Anti-hnRNP A2B1 antibody (ab31645)
Key features and details
- Rabbit polyclonal to hnRNP A2B1
- Suitable for: IHC-P, WB, ICC/IF, IP
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-hnRNP A2B1 antibody
See all hnRNP A2B1 primary antibodies -
Description
Rabbit polyclonal to hnRNP A2B1 -
Host species
Rabbit -
Specificity
The peptide used as the immunogen for this antibody is found within isoform A2 and B1 of the protein and should thus recognize a doublet around 35/37 kDa. -
Tested Applications & Species
Application Species ICC/IF HumanIHC-P HumanIP HumanWB Human -
Immunogen
Synthetic peptide corresponding to Human hnRNP A2B1 aa 50-150.
(Peptide available asab31644) -
Positive control
- WB: A549, HEK293T, HeLa, NIH 3T3, and MEF1 cell lysates, and mouse testis tissue lysate. IHC-P: Human colon tissue.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab31645 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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ICC/IF |
Human
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IHC-P |
Human
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IP |
Human
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WB |
Human
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All applications |
Mouse
Rat
Chicken
Dog
Xenopus laevis
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Application | Abreviews | Notes |
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IHC-P |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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WB | (2) |
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 35, 38 kDa (predicted molecular weight: 35 , 37 kDa).
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ICC/IF | (3) |
Use a concentration of 5 µg/ml.
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IP |
Use at an assay dependent concentration.
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Notes |
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IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 35, 38 kDa (predicted molecular weight: 35 , 37 kDa). |
ICC/IF
Use a concentration of 5 µg/ml. |
IP
Use at an assay dependent concentration. |
Target
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Function
Involved with pre-mRNA processing. Forms complexes (ribonucleosomes) with at least 20 other different hnRNP and heterogeneous nuclear RNA in the nucleous. -
Sequence similarities
Contains 2 RRM (RNA recognition motif) domains. -
Cellular localization
Nucleus > nucleoplasm. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Component of ribonucleosomes. Predominantly nucleoplasmic, however isoform A2 is also found in the cytoplasm of cells in some tissues. Not found in the nucleolus. - Information by UniProt
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Database links
- Entrez Gene: 3181 Human
- Entrez Gene: 100045191 Mouse
- Entrez Gene: 53379 Mouse
- Entrez Gene: 362361 Rat
- Entrez Gene: 380082 Xenopus laevis
- Omim: 600124 Human
- SwissProt: P22626 Human
- SwissProt: O88569 Mouse
see all -
Alternative names
- Heterogeneous nuclear ribonucleoprotein A2 antibody
- Heterogeneous nuclear ribonucleoprotein A2/B1 antibody
- Heterogeneous nuclear ribonucleoprotein B1 antibody
see all
Images
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All lanes : Anti-hnRNP A2B1 antibody (ab31645) at 1/500 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : HNRNPA2B1 knockout HEK293T cell lysate
Lane 3 : A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 35 , 37 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab31645 observed at 37 kDa. Red - loading control, ab7291 observed at 50 kDa.
ab31645 Anti-hnRNP A2B1 antibody was shown to specifically react with hnRNP A2B1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266404 (knockout cell lysate ab257224) was used. Wild-type and hnRNP A2B1 knockout samples were subjected to SDS-PAGE. ab31645 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti- Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti- Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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Anti-hnRNP A2B1 antibody (ab31645) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 35 , 37 kDa
Observed band size: 35,38 kDa why is the actual band size different from the predicted?
ab31645 is expected to recognize isoform hnrnp A2 and hnrnp B1. The antibody detects bands at approximately 35 and 38 kDa which are of the correct size to correspond to the predicted molecular weight of these isoforms. -
Immunocytochemistry/ Immunofluorescence - Anti-hnRNP A2B1 antibody (ab31645)This image is courtesy of an anonymous Abreview
ab31645 staining hnRNP A2B1 in CHO cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 1 hour at 21°C. Samples were incubated with primary antibody (1/200 in PBS + 1% BSA) for 1 hour 30 minutes at 21°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody. Green - hnRNP, Red -alpha tubulin, Blue - nuclei.
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hnRNP A2B1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to hnRNP A2B1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31645.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 35kDa: hnRNP A2B1. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP A2B1 antibody (ab31645)IHC image of ab31645 staining in human colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab31645, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
ICC/IF image of ab31645 stained human MCF7 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab31645, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293 and HepG2 cells at 5µg/ml.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (14)
ab31645 has been referenced in 14 publications.
- Han M et al. Exosome-mediated lncRNA AFAP1-AS1 promotes trastuzumab resistance through binding with AUF1 and activating ERBB2 translation. Mol Cancer 19:26 (2020). PubMed: 32020881
- Wiktorowicz JE et al. Integrated Functional Analysis of the Nuclear Proteome of Classically and Alternatively Activated Macrophages. Mediators Inflamm 2019:3481430 (2019). PubMed: 31182931
- Koppers M et al. Receptor-specific interactome as a hub for rapid cue-induced selective translation in axons. Elife 8:N/A (2019). PubMed: 31746735
- Liu Y et al. The UCA1/KRAS axis promotes human pancreatic ductal adenocarcinoma stem cell properties and tumor growth. Am J Cancer Res 9:496-510 (2019). PubMed: 30949406
- Zheng Z et al. Increased Expression of Exosomal AGAP2-AS1 (AGAP2 Antisense RNA 1) In Breast Cancer Cells Inhibits Trastuzumab-Induced Cell Cytotoxicity. Med Sci Monit 25:2211-2220 (2019). PubMed: 30910994