Immunofluorescence (IF)
(Immunofluorescence staining of fixed HeLa cells with anti-H4 K8Ac; K12Ac; K16Ac antibody KM-2 (MBS488786) Immunofluorescence analysis of paraformaldehyde fixed HeLa cells on Shi-fix coverslips stained with the chimeric rabbit IgG version of KM-2 (MBS488786) at 10ug/ml for 1h followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing membrane staining. The nuclear stain is DAPI (blue) and the actin stain is phalloidin (red). Panels show from left-right, top-bottom MBS488786, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (MBS488149) followed by staining with Alexa Fluor 488 secondary antibody.)
Mouse anti-Human H4 K8Ac; K12Ac; K16Ac Monoclonal Antibody
Anti-H4 K8Ac; K12Ac; K16Ac [KM-2]
Reactivity
Human
Applications
ELISA, Western Blot, Immunofluorescence, Immunohistochemistry
Purity
Purified antibody. Protein A affinity purified
Synonyms
H4 K8Ac; K12Ac; K16Ac; Monoclonal Antibody; Anti-H4 K8Ac; K16Ac [KM-2]; histone 4; nucleosome; H4K8Ac; H4K12Ac; H4K16Ac; acetyl-Histone H4; Histone H4 acetylated at lysine; KM2; anti-H4 K8Ac; K16Ac antibody
Host
Mouse
Reactivity
Human
Clonality
Monoclonal
Isotype
Mouse IgG2a, kappa
Clone Number
KM-2
Specificity
KM-2 antibody recognizes the N-terminal peptides 1-20 of histone H2A and 1-29 of histone H4 which have a large sequence homology (van Bruggen et al., 1997; pmid: 9027774). Histone H4 acetylation on lysines 8, 12 and 16 significantly increases the antibody's binding while the acetylation at K5 decreases it (Dieker et al., 2007; pmid: 17530637).
Purity/Purification
Purified antibody. Protein A affinity purified
Form/Format
PBS with 0.02% Proclin 300.
Concentration
1mg/ml (varies by lot)
Applicable Applications for anti-H4 K8Ac; K12Ac; K16Ac antibody
ELISA (EIA), Western Blot (WB), Immunofluorescence (IF), Immunohistochemistry (IHC)
Application Notes
The antibody KM-2 shows significant increase in activity with histone H4 when it is acethylated on lysines 8, 12 and 16 which are hyperacethylated in conditions, such as lupus (SLE); and thus KM-2 can be used in the detection of the apoptosis-induced histone modifications characteristic of lupus and in the analysis of its pathogenesis (Dieker et al., 2007; pmid: 17530637). This antibody was successfully used to identify both Histone H4 as well as H2A in the extracts of Jurkat cells via Western blot, showing considerable increase in activity in cells with induced apoptosis leading to hyperacethylation (Dieker et al., 2007; pmid: 17530637). Similar results were achieved for the immunofluorescence staining of Jurkat cells. The enhanced reactivity of KM?2 with apoptotic H4 and apoptotic H2A was not Jurkat cell specific and was also observed for the monocytic U937 cell line which indicated its general applicability (Dieker et al., 2007; pmid: 17530637). Analogous reselts were also achieved during the ELISA analysis of plasma samples of (diseased) MRL/lpr lupus mice, with the visisble activity enhancement in the sick, apoptotic specimens (Dieker et al., 2007; pmid: 17530637). Finally, KM-2 (KM2) antibody was used to identify histones in the immune deposits in kidney biopsy samples of DPGN (Diffuse proliferative glomerulonephritis) patients (van Bruggen et al., 1997; pmid: 9027774).
PMID
9027774
Preparation and Storage
Store at 4 degree C for up to 3 months. For longer storage, aliquot and store at -20 degree C.
Immunofluorescence (IF)
(Immunofluorescence staining of fixed HeLa cells with anti-H4 K8Ac; K12Ac; K16Ac antibody KM-2 (MBS488786) Immunofluorescence analysis of paraformaldehyde fixed HeLa cells on Shi-fix coverslips stained with the chimeric rabbit IgG version of KM-2 (MBS488786) at 10ug/ml for 1h followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing membrane staining. The nuclear stain is DAPI (blue) and the actin stain is phalloidin (red). Panels show from left-right, top-bottom MBS488786, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (MBS488149) followed by staining with Alexa Fluor 488 secondary antibody.)
Immunofluorescence (IF)
(Immunofluorescence staining of fixed HeLa cells with anti-H4 K8Ac; K12Ac; K16Ac antibody KM-2 (MBS488786) Immunofluorescence analysis of paraformaldehyde fixed HeLa cells on Shi-fix coverslips stained with the chimeric rabbit IgG version of KM-2 (MBS488786) at 10ug/ml for 1h followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing membrane staining. The nuclear stain is DAPI (blue) and the actin stain is phalloidin (red). Panels show from left-right, top-bottom MBS488786, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (MBS488149) followed by staining with Alexa Fluor 488 secondary antibody.)
Flow Cytometry (FC/FACS)
(Flow cytometry using the anti-H4 K8Ac; K12Ac; K16Ac antibody KM-2 (MBS488786). HeLa cells were fixed using 2% PFA and stained with anti-unknown specificity antibody (MBS488149; isotype control, black line) or the rabbit IgG1 version of KM-2 (MBS488786, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.)
Flow Cytometry (FC/FACS)
(Flow cytometry using the anti-H4 K8Ac; K12Ac; K16Ac antibody KM-2 (MBS488786). HeLa cells were fixed using 2% PFA and stained with anti-unknown specificity antibody (MBS488149; isotype control, black line) or the rabbit IgG1 version of KM-2 (MBS488786, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.)
Related Product Information for anti-H4 K8Ac; K12Ac; K16Ac antibody
Recombinant monoclonal antibody to H4 K8Ac; K12Ac; K16Ac. Recombinant Platform with variable regions (i.e. specificity) from the hybridoma KM-2.
KM-2 antibody was derived from an MRL/lpr lupus mouse.
KM-2 antibody was derived from an MRL/lpr lupus mouse.
Product Categories/Family for anti-H4 K8Ac; K12Ac; K16Ac antibody
References
van Bruggen et al. Nucleosomes and histones are present in glomerular deposits in human lupus nephritis. Nephrol Dial Transplant. 1997 Jan;12(1):57-66.
Note on publication: This article describes the characterization of the KM-2 (KM2) antibody.
Note on publication: This article describes the characterization of the KM-2 (KM2) antibody.
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Product Notes
The H4 K8Ac; K12Ac; K16Ac (Catalog #AAA488785) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-H4 K8Ac; K12Ac; K16Ac [KM-2] reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's H4 K8Ac; K12Ac; K16Ac can be used in a range of immunoassay formats including, but not limited to, ELISA (EIA), Western Blot (WB), Immunofluorescence (IF), Immunohistochemistry (IHC). The antibody KM-2 shows significant increase in activity with histone H4 when it is acethylated on lysines 8, 12 and 16 which are hyperacethylated in conditions, such as lupus (SLE); and thus KM-2 can be used in the detection of the apoptosis-induced histone modifications characteristic of lupus and in the analysis of its pathogenesis (Dieker et al., 2007; pmid: 17530637). This antibody was successfully used to identify both Histone H4 as well as H2A in the extracts of Jurkat cells via Western blot, showing considerable increase in activity in cells with induced apoptosis leading to hyperacethylation (Dieker et al., 2007; pmid: 17530637). Similar results were achieved for the immunofluorescence staining of Jurkat cells. The enhanced reactivity of KM?2 with apoptotic H4 and apoptotic H2A was not Jurkat cell specific and was also observed for the monocytic U937 cell line which indicated its general applicability (Dieker et al., 2007; pmid: 17530637). Analogous reselts were also achieved during the ELISA analysis of plasma samples of (diseased) MRL/lpr lupus mice, with the visisble activity enhancement in the sick, apoptotic specimens (Dieker et al., 2007; pmid: 17530637). Finally, KM-2 (KM2) antibody was used to identify histones in the immune deposits in kidney biopsy samples of DPGN (Diffuse proliferative glomerulonephritis) patients (van Bruggen et al., 1997; pmid: 9027774). Researchers should empirically determine the suitability of the H4 K8Ac; K12Ac; K16Ac for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "H4 K8Ac; K12Ac; K16Ac, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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