Overview

  • Product name
  • Description
    Rabbit polyclonal to hnRNP K
  • Specificity
    This antibody strongly detects a single band at ~65 kDa. The predicted molecular weight of hnRNP K is ~51 kDa, but it has been shown to migrate around 65 kDa (see Milak and Clements, Nucleic Acids Research, vol 32(18), pp 5553-5569). This band is competed away by the addition of the immunizing peptide.
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Rabbit, Chicken, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human hnRNP K.

    (Peptide available as ab19120.)

  • Positive control
    • ab18195 gave a positive result in Mouse Testis and Ovary Tissue Lysates, HeLa nuclear extract and the following whole cell lysates: HeLa; Jurkat; A431; 293T; NIH3T3; MEF1; PC12. This antibody gave a positive result in IHC in the following FFPE tissue: Human Colon Adenocarcinoma.

Properties

  • 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
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab18195 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 51 kDa).

Target

Images

  • IHC image of hnRNP K staining in Human Colon Adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab18195, 0.5 µg/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 ab18195 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab18195, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

    Panel A (left) shows localisation of the antibody (green) to specific foci in the nucleus, panel B (right) does not show the Alexa Fluor® 488 channel for comparison.

  • All lanes : Anti-hnRNP K antibody (ab18195) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line)
    Lane 3 : Testis (Mouse) Tissue Lysate
    Lane 4 : Ovary (Mouse) Tissue Lysate
    Lane 5 : PC12 (Rat adrenal pheochromocytoma cell line)

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Predicted band size : 51 kDa
    Observed band size : 65 kDa (why is the actual band size different from the predicted?)
  • ab18195 staining hnRNP K in HeLa cells treated with PD 98059 (ab120234), by ICC/IF. Changes in localization of hnRNP K (translocation from cytoplasm to nucleous) correlates with increased concentration of hnRNP K, as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120234 ( hnRNP K) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab18195(5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

References

This product has been referenced in:
  • Moujalled D  et al. Kinase Inhibitor Screening Identifies Cyclin-Dependent Kinases and Glycogen Synthase Kinase 3 as Potential Modulators of TDP-43 Cytosolic Accumulation during Cell Stress. PLoS One 8:e67433 (2013). ICC/IF ; Human . Read more (PubMed: 23840699) »
  • van Domselaar R  et al. Granzyme M targets host cell hnRNP K that is essential for human cytomegalovirus replication. Cell Death Differ : (2012). Read more (PubMed: 23099853) »

See all 5 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
CLIP
Sample
Human Cell lysate - whole cell (HEK 293T)
Specification
HEK 293T
Type
iCLIP
Irradiation Energy Level (mJ/cm2): 150
Wavelength (nm): 254
Positive control
hnRNP C antibody (sc-15386) bound protein A dynabeads 0.1 mg/ml (antibody/beads).
Negative control
Protein A dynabeads without antibody bound.
Immuno-precipitation step
Other - Protein A dynabeads
Username

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Verified customer

Submitted Aug 18 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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