Anti-hnRNP K antibody (ab32969)
Key features and details
- Rabbit polyclonal to hnRNP K
- Suitable for: IP, WB, ICC/IF, IHC-P
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-hnRNP K antibody
See all hnRNP K primary antibodies -
Description
Rabbit polyclonal to hnRNP K -
Host species
Rabbit -
Tested applications
Suitable for: IP, WB, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Rabbit, Chicken, Zebrafish
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Immunogen
Synthetic peptide corresponding to Human hnRNP K aa 450 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available asab32584) -
Positive control
- This antibody gave positive signal in the following whole cell lysates: HeLa; A431; HepG2; U2OS; NIH 3T3; MEF1; PC12. This antibody gave positive signal in the following Mouse tissue lysates: Brain; Testis; Ovary. This antibody gave a positive result in IHC in the following FFPE tissue: Human Colon Adenocarcinoma.
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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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Images
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Western blot - Anti-hnRNP K antibody (ab32969)All lanes : Anti-hnRNP K antibody (ab32969) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : Hep G2 whole cell lysate (ab7900)
Lane 4 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution
Performed under reducing conditions.
Predicted band size: 51 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP K antibody (ab32969)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 ab32969, 1 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.
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Immunoprecipitation - Anti-hnRNP K antibody (ab32969)hnRNP K was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to hnRNP K 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 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32969.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 50kDa; hnRNP K
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Western blot - Anti-hnRNP K antibody (ab32969)All lanes : Anti-hnRNP K antibody (ab32969) at 1 µg/ml
Lane 1 :NIH/3T3 whole cell lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : Brain (Mouse) Tissue Lysate
Lane 4 : Testis (Mouse) Tissue Lysate - normal tissue
Lane 5 : Ovary (Mouse) Tissue Lysate - normal tissue
Lane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 7 : Brain (Rat) Tissue Lysate - normal tissue
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 51 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted? -
Immunocytochemistry/ Immunofluorescence - Anti-hnRNP K antibody (ab32969)ICC/IF image of ab32969 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab32969, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and 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).
