Mouse Recombinant Monoclonal H2B antibody. Suitable for IHC-P, IP, ChIP, Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse, Rat, Recombinant full length protein - Human samples. Cited in 45 publications.
IgG1
Mouse
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IHC-P | IP | ChIP | Flow Cyt (Intra) | WB | ICC/IF | |
---|---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested | Tested |
Mouse | Tested | Tested | Expected | Tested | Tested | Tested |
Rat | Tested | Expected | Expected | Expected | Tested | Expected |
Caenorhabditis elegans | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Chicken | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Cow | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Drosophila melanogaster | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Orangutan | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Recombinant full length protein - Human | Not recommended | Not recommended | Not recommended | Not recommended | Expected | Not recommended |
Xenopus laevis | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Zebrafish | Predicted | Predicted | Predicted | Predicted | Predicted | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/20000 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Mouse | Dilution info 1/50000 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/50000 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Chicken, Cow, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish, Orangutan | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant full length protein - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes - |
Species Mouse | Dilution info 1/50 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Chicken, Cow, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish, Orangutan | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant full length protein - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 2 µg for 25 µL chromatin | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Chicken, Cow, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish, Orangutan | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant full length protein - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Chicken, Cow, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish, Orangutan | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant full length protein - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes - |
Species Mouse | Dilution info 1.00000-5.00000 µg/mL | Notes - |
Species Rat | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant full length protein - Human | Dilution info 1.00000-5.00000 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Chicken, Cow, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish, Orangutan | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/2000 | Notes - |
Species Mouse | Dilution info 1/2000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Chicken, Cow, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish, Orangutan | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant full length protein - Human | Dilution info - | Notes - |
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Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
H2BFF, HIST1H2BB, HIST1H2BB, H2BFF, H2BC3, Histone H2B type 1-B, H2B-clustered histone 3, Histone H2B.1, Histone H2B.f, H2B/f
Mouse Recombinant Monoclonal H2B antibody. Suitable for IHC-P, IP, ChIP, Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse, Rat, Recombinant full length protein - Human samples. Cited in 45 publications.
IgG1
Mouse
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
mAbcam 52484
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25μg of chromatin, 5μg of ab52484 (blue), and 20μl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
This image was generated using a previous batch manufactured using hybridoma production method.
ab52484 staining Histone H2B in Fruit fly (Drosophila melanogaster) embryo cells by ICC/IF (Immunocytochemistry/immunofluorescence). Embryos were washed in 1x PBS + 0.1% Trition, cells were fixed with heat, and blocked with 10% BSA for 12 hours at 4°C. Samples were incubated with primary antibody (1/1000) for 24 hours. An Alexa Fluor®594-conjugated Rabbit anti-mouse IgG polyclonal (1/5000) was used as the secondary antibody.
Histone H2B was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Histone H2B 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 ab52484.
Secondary: Protein G-HRP at 1/500 dilution.
Band: 17kDa; Histone H2B; non specific bands - 26kDa: We are unsure as to the identity of this extra band.
All lanes: Immunoprecipitation - Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484)
Predicted band size: 14 kDa
This image was generated using a
previous batch manufactured using hybridoma production method
All lanes: Western blot - Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 5 µg/mL
Lane 1: Marker
Lane 2: Zebrafish brain homogenate at 20 µg
Lane 3: Zebrafish heart homogenate at 10 µg
Lane 4: Zebrafish liver homogenate at 10 µg
Lane 5: Zebrafish skeletal muscle homogenate at 10 µg
Lane 6: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
All lanes: Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 17 kDa
Exposure time: 1min
All lanes: Western blot - Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 5 µg/mL
Lane 1: Histone H1 recombinant protein at 0.1 µg
Lane 2: Histone H2A recombinant protein at 0.1 µg
Lane 3: Histone H2B recombinant protein at 0.1 µg
Lane 4: Histone H3 recombinant protein at 0.1 µg
Lane 5: Histone H4 recombinant protein at 0.1 µg
All lanes: Rabbit polyclonal to Mouse IgG - H&L (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 17 kDa
ICC/IF image of ab52484 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab52484, 1μg/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).
IHC image of Histone H2B staining in human breast carcinoma FFPE section, performed on a BondTM 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 ab52484, 1µg/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.
All lanes: Western blot - Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 1 µg/mL
Lane 1: NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg
Lane 2: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg
All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (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: 14 kDa
Observed band size: 17 kDa
Exposure time: 1min
This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arignine, and sodium azide (ab52484).
Overlay histogram showing HeLa cells stained with ab52484 (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 (ab52484, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
All lanes: Western blot - Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 1/1000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 2: 293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 3: MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate at 20 µg
Lane 4: PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg
Lane 5: Human heart tissue lysate at 20 µg
Lane 6: Mouse heart tissue lysate at 20 µg
All lanes: Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution
Predicted band size: 14 kDa
Observed band size: 15 kDa
Exposure time: 3.25s
Histone H2B was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast), whole cell lysate with ab52484 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab52484 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 μg
Lane 2: ab52484 IP in NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 μg
Lane 3: Mouse monoclonal IgG (Mouse IgG1, kappa monoclonal [MOPC-21] - isotype control ab18443) instead of ab52484 in NIH/3T3 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds.
All lanes: Immunoprecipitation - Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 1/30 dilution
All lanes: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 2 µg of ab52484 (red), or 2 µg of mouse IgG1 Mouse IgG1, kappa monoclonal [MOPC-21] - isotype control ab18443 (gray) and 25 µl of Protein A/G Dynabeads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Histone H2B was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab52484 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab52484 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug
Lane 2: ab52484 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3: Mouse monoclonal IgG (Mouse IgG1, kappa monoclonal [MOPC-21] - isotype control ab18443) instead of ab52484 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds.
All lanes: Immunoprecipitation - Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade (ab52484) at 1/30 dilution
All lanes: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Predicted band size: 14 kDa
Observed band size: 15 kDa
Flow cytometric analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Histone H2B with ab52484 at 1/1000 dilution (0.1µg) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti mouse IgG (Alexa Fluor® 488, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/2000 dilution was used as the secondary antibody.
Flow cytometric analysis of NIH/3T3 (mouse embryonic fibroblast) cells labelling Histone H2B with ab52484 at 1/100 dilution (1µg) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti mouse IgG (Alexa Fluor® 488, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/2000 dilution was used as the secondary antibody.
NIH/3T3 cells were fixed in 100% methanol and permeabilized with 0.1% Triton X-100. Primary antibody, ab52484 at 1:2000 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with ab52484 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
HeLa cells were fixed in 100% methanol and permeabilized with 0.1% Triton X-100. Primary antibody, ab52484 at 1:2000 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with ab52484 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
Nuclear staining in rat lung. The section was heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The section was incubated with ab52484 (1:50000 (0.021 μg/ml)) for 10 mins at room temperature, followed by anti-mouse IgG1 antibody (Rabbit monoclonal [M1gG51-4] Anti-Mouse IgG1 H&L ab125913) for 8 mins during the LeicaDS9800 kit staining procedure.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Nuclear staining in mouse lung. The section was heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The section was incubated with ab52484 (1:50000 (0.021 μg/ml)) for 10 mins at room temperature, followed by anti-mouse IgG1 antibody (Rabbit monoclonal [M1gG51-4] Anti-Mouse IgG1 H&L ab125913) for 8 mins during the LeicaDS9800 kit staining procedure.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Nuclear staining in human breast cancer. The section was heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
The section was incubated with ab52484 at 1:20000 (0.053 μg/ml) for 10 mins at room temperature, followed by anti-mouse IgG1 antibody (Rabbit monoclonal [M1gG51-4] Anti-Mouse IgG1 H&L ab125913) for 8 mins during the LeicaDS9800 kit staining procedure.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
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