Anti-hnRNP K antibody [F45 P9 C7] (ab23644)

Reviews (2) Submit a question References (9)

Overview

  • Product name

    Anti-hnRNP K antibody [F45 P9 C7]
    See all hnRNP K primary antibodies

  • Description

    Mouse monoclonal [F45 P9 C7] to hnRNP K

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Rabbit, Chicken

  • Immunogen

    Synthetic peptide corresponding to Human hnRNP K aa 450 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab32584)

  • Positive control

    • This antibody gave a positive signal in the following lysates: NIH 3T3 whole cell, MEF1 whole cell; PC12 whole cell; mouse testis tissue; mouse lung tissue.

  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

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

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Some batches contain 6.97% L-Arginine as a stabilizing agent. For lot-specific buffer information, please contact our Scientific Support team.
  • Concentration information loading...

  • Purity

    IgG fraction

  • Clonality

    Monoclonal

  • Clone number

    F45 P9 C7

  • Myeloma

    Ag8.653

  • Isotype

    IgG1

  • Light chain type

    kappa

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab23644 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 5 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
WB Use a concentration of 1 µg/ml. Detects a band of approximately 64 kDa (predicted molecular weight: 51 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.

Target

  • Function

    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.

  • Sequence similarities

    Contains 3 KH domains.

  • Post-translational
    modifications

    Arg-296 and Arg-299 are dimethylated, probably to asymmetric dimethylarginine.

  • Cellular localization

    Cytoplasm. Nucleus > nucleoplasm. In case of ASFV infection, there is a shift in the localization which becomes predominantly nuclear.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • CSBP antibody
    • dC stretch binding protein antibody
    • FLJ41122 antibody
    • Heterogeneous nuclear ribonucleoprotein K antibody
    • hnRNP K antibody
    • HNRNPK antibody
    • HNRPK antibody
    • HNRPK_HUMAN antibody
    • Transformation up regulated nuclear protein antibody
    • Transformation up-regulated nuclear protein antibody
    • Transformation upregulated nuclear protein antibody
    • TUNP antibody
    see all

Images

  • Western blot - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    Western blot - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    Lane 1 : Marker
    Lanes 2-5 : Anti-hnRNP K antibody [F45 P9 C7] (ab23644) at 1 µg/ml

    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate (ab27251) at 20 µg
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 20 µg
    Lane 4 : Jurkat whole cell lysate (ab7899) at 20 µg
    Lane 5 : HEK293 whole cell lysate (ab7902) at 20 µg

    Secondary
    Lanes 2-5 : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 51 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    Formalin-fixed, paraffin embedded tissue sections from human normal colon or human colon carcinoma stained with ab23644 mouse monoclonal to hnRNP K. hnRNP K expression is increased in the tumour tissue.
  • Immunocytochemistry/ Immunofluorescence - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    Immunocytochemistry/ Immunofluorescence - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)

    ICC/IF image of ab23644 stained human HepG2 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab23644, 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-mouse 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 IF result in HeLa, HEK 293 and MCF7 cells.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)This image is courtesy of an Abreview submitted by Satyendra Tripathi
    ab23644 staining hnRNP K in Human head and neck tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 1 hour at 25°C; antigen retrieval was by heat mediation with Tris EDTA (pH9). Samples were incubated with primary antibody (1 µg/ml in TBST) for 16 hours at 4°C. An undiluted HRP-conjugated Rabbit anti-mouse polyclonal was used as the secondary antibody.

    See Abreview

  • Western blot - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    Western blot - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    All lanes : Anti-hnRNP K antibody [F45 P9 C7] (ab23644) at 5 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Testis (Mouse) Tissue Lysate
    Lane 4 : Lung (Mouse) Tissue Lysate
    Lane 5 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 51 kDa
    Observed band size: 56 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 21 kDa, 24 kDa, 54 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes
  • Flow Cytometry - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    Flow Cytometry - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    Overlay histogram showing HeLa cells stained with ab23644 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab23644, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Immunocytochemistry/ Immunofluorescence - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)
    Immunocytochemistry/ Immunofluorescence - Anti-hnRNP K antibody [F45 P9 C7] (ab23644)Image from Meyerowitz J et al., Mol Neurodegener. 2011 Aug 8;6:57. Fig 10.; doi:10.1186/1750-1326-6-57; 8 August 2011, Molecular Neurodegeneration 2011, 6:57
    Immunofluorescence analysis of SH-SY5Y cells staining hnRNP K with ab23644.

    Cells were fixed with paraformaldehyde in PBS for 30 min and permeabilized with 90% chilled methanol for 5 min. After blocking for 1 hour with 10% normal goat serum, cells were incubated with primary antibody (1/200) overnight at 4°C. An AlexaFluor®-conjugated goat anti-mouse IgG (1/500) was used as the secondary antibody.

References

This product has been referenced in:

  • Zhao S  et al. hnRNP K plays a protective role in TNF-a-induced apoptosis in podocytes. Biosci Rep 38:N/A (2018). WB . Read more (PubMed: 29724888) »
  • Pellegrini L  et al. Proteomic analysis reveals co-ordinated alterations in protein synthesis and degradation pathways in LRRK2 knockout mice. Hum Mol Genet 27:3257-3271 (2018). Read more (PubMed: 29917075) »
See all 9 Publications for this product

Publishing research using ab23644? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-2 of 2 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-hnRNP K antibody [F45 P9 C7]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Brain Frontal Cortex)
Specification
Brain Frontal Cortex
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Acetone
Read More

Abcam user community

Verified customer

Submitted Jan 21 2020

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-hnRNP K antibody [F45 P9 C7]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Head and neck)
Specification
Head and neck
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9.0
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More

Dr. Satyendra Tripathi

Verified customer

Submitted Sep 14 2010

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com