Anti-HMG14 antibody - ChIP Grade
| Name | Anti-HMG14 antibody - ChIP Grade |
|---|---|
| Supplier | Abcam |
| Catalog | ab5212 |
| Prices | $401.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | ICC/IF ICC/IF ChIP ICC/IF IP IHC-P FC WB |
| Species Reactivities | Mouse, Rat, Human, Bovine |
| Antigen | Synthetic peptide derived from residues 50 to the C-terminus of Human HMGN1/ HMG14 |
| Blocking Peptide | Human HMG14 peptide |
| Description | Rabbit Polyclonal |
| Gene | HMGN1 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
Ab5212 stained HeLa cells. The cells were 4% PFA fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab5212 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Confocal microscopy of fixed primary cultures of rat (embryonic day 18) hippocampal neurons showing immunocytochemical labelling of monoclonal mouse anti-beta-tubulin-II (1/400; Alexa Fluor 568; red) and polyclonal rabbit anti-HMGN1 (ab5212, 1/500; Alexa Fluor 488; green). Magnification 63x ; insert 240x. Immunofluorescence revealed exclusive nuclear staining in rat cultured cells, corresponding to the known nuclear localisation of HMG14.
Fluorescence microscopy of fixed murine neurospheres showing (left to right) immunocytochemical labeling with polyclonal rabbit anti-HMG14 (ab5212 at 1/500; Alexa Fluor 488; green), and monoclonal mouse anti-nestin (1/500; Alexa Fluor 568; red) with their overlay (far right). Scale bar = 30µm.
Image courtesy of Human Protein Atlas ab5212 staining HMG14 in Human skin, showing a distinct and strong staining pattern of the epidermal cells. Paraffin embedded human skin tissue was incubated with ab5212 (1/3000) for 30 minutes at room temperature.Antigen retrieval was performed by heat induction in citrate buffer pH6. ab5212 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
Chromatin was prepared from the Human embryonic stem cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. The primary antibody was diluted 1/100 in IP dilution buffer and incubated with the sample for 12 hours at 4°C. Histone H3 was used as the positive control, whilst rabbit IgG was used as the negative control. The immunoprecipitated DNA was quantified by real time PCR.See Abreview
HMG14 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to HMG14 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 ab5212.Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.Band: 17kDa: HMG14
Performed under reducing conditions.
Performed under reducing conditions.
Performed under reducing conditions.
Performed under reducing conditions.
Performed under reducing conditions.
ab5212 staining HMG14 in human SH-SY5Y cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized. The sample was incubated with the primary antibody (1/500) for 1 hour at 20°C. A PerCP-conjugated goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.Gating Strategy: isotype population (shown in white).See Abreview
Immunofluorescent imaging of human cells (U2OS) with ab5212 reveals the expected exclusively nuclear staining, corresponding to the known nuclear localisation of HMGN1/HMG14.IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour. Primary antibody used at 5ug/ml in 5% milk / 0.2% TWEEN for one hour, secondary antibody for 30 minutes. All blocking and incubation steps carried out at 37 degrees C. Nuclear counterstaining with Hoechst stain (blue).
Product References
Triplication of a 21q22 region contributes to B cell transformation through HMGN1 - Triplication of a 21q22 region contributes to B cell transformation through HMGN1
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Reprogramming ovarian and breast cancer cells into non-cancerous cells by - Reprogramming ovarian and breast cancer cells into non-cancerous cells by
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Transcription-dependent dynamic supercoiling is a short-range genomic force. - Transcription-dependent dynamic supercoiling is a short-range genomic force.
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UV-sensitive syndrome protein UVSSA recruits USP7 to regulate - UV-sensitive syndrome protein UVSSA recruits USP7 to regulate
Schwertman P, Lagarou A, Dekkers DH, Raams A, van der Hoek AC, Laffeber C, Hoeijmakers JH, Demmers JA, Fousteri M, Vermeulen W, Marteijn JA. Nat Genet. 2012 May;44(5):598-602.
Identification of nuclear protein targets for six leukemogenic tyrosine kinases - Identification of nuclear protein targets for six leukemogenic tyrosine kinases
Pierce A, Williamson A, Jaworska E, Griffiths JR, Taylor S, Walker M, Aspinall-O'Dea M, Spooncer E, Unwin RD, Poolman T, Ray D, Whetton AD. PLoS One. 2012;7(6):e38928.
FOXO3 signalling links ATM to the p53 apoptotic pathway following DNA damage. - FOXO3 signalling links ATM to the p53 apoptotic pathway following DNA damage.
Chung YM, Park SH, Tsai WB, Wang SY, Ikeda MA, Berek JS, Chen DJ, Hu MC. Nat Commun. 2012;3:1000.
Dynamic changes in histone H3 phosphoacetylation during early embryonic stem cell - Dynamic changes in histone H3 phosphoacetylation during early embryonic stem cell
Lee ER, McCool KW, Murdoch FE, Fritsch MK. J Biol Chem. 2006 Jul 28;281(30):21162-72. Epub 2006 May 25.