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.
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.
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.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 51 kDa).
One of the major pre-mRNA-binding proteins. Binds tenaciously to poly(C) sequences. Likely to play a role in the nuclear metabolism of hnRNAs, particularly for pre-mRNAs that contain cytidine-rich sequences. Can also bind poly(C) single-stranded DNA.
Contains 3 KH domains.
Arg-296 and Arg-299 are dimethylated, probably to asymmetric dimethylarginine.
Cytoplasm. Nucleus > nucleoplasm. In case of ASFV infection, there is a shift in the localization which becomes predominantly nuclear.
Heterogeneous nuclear ribonucleoprotein K antibody
hnRNP K antibody
Transformation up regulated nuclear protein antibody
Transformation up-regulated nuclear protein antibody
Transformation upregulated nuclear protein antibody
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP K antibody (ab18195)
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.
Immunocytochemistry/ Immunofluorescence - hnRNP K antibody (ab18195)
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.
Western blot - hnRNP K antibody (ab18195)
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)
Immunocytochemistry/ Immunofluorescence-Anti-hnRNP K antibody(ab18195)
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.
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 One8:e67433 (2013).
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) »