Anti-HDAC1 antibody [EPR23847-170] (ab280198)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23847-170] to HDAC1
- Suitable for: Flow Cyt (Intra), ChIP, WB, IHC-P, ICC, IP
- Reacts with: Mouse, Rat, Human
Overview
-
Product name
Anti-HDAC1 antibody [EPR23847-170]
See all HDAC1 primary antibodies -
Description
Rabbit monoclonal [EPR23847-170] to HDAC1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ChIP MouseFlow Cyt MouseICC HumanWB MouseRatHumanRecombinant fragment -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Human heart and kidney tissue lysates; Jurkat, K-562, HeLa, C6, RAW264.7, PC-12 and NIH/3T3 whole cell lysates; His-tagged human HDAC1 recombinant protein. IHC-P: Human tonsil and liver tissue; Mouse liver tissue; Rat liver tissue. ICC: HeLa and NIH/3T3 cells. Flow Cyt: HeLa cells and NIH/3T3 cells. IP: HeLa and NIH/3T3 whole cell lysates. ChIP: Chromatin prepared from K-562 cells.
-
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23847-170 -
Isotype
IgG -
Research areas
Images
-
Chromatin was prepared from K-562 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab280198 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 25 µl of Protein A/G Dynabeads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).*https://www.abcam.com/resources?keywords=X%20ChIP%20protocol
-
HDAC1 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab280198 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280198 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug
Lane 2: ab280198 IP in NIH/3T3 whole cell lysate 10 ug
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab280198 in NIH/3T3 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
-
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labelling HDAC1 with ab280198 at 1/5000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear and weakly cytoplasmic staining in HeLa cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR23847-170] (ab280198)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling HDAC1 with ab280198 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human tonsil (PMID:23109994). The section was incubated with ab280198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
All lanes : Anti-HDAC1 antibody [EPR23847-170] (ab280198) at 1/1000 dilution
Lane 1 : Human heart tissue lysate
Lane 2 : Human kidney tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution (VeriBlot for IP secondary antibody(HRP))
Predicted band size: 55 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID:24551070).
Exposure time: 15 seconds.
-
Flow cytometric analysis of 4% paraformaldehyde-fixe,d 90% methanol-permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling HDAC1 with ab280198 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
-
All lanes : Anti-HDAC1 antibody [EPR23847-170] (ab280198) at 1/5000 dilution
Lane 1 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
Lane 2 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 55 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID: 24551070).
Exposure time: 37 seconds.
-
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labelling HDAC1 with ab280198 at 1/5000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear and weakly cytoplasmic staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
-
All lanes : Anti-HDAC1 antibody [EPR23847-170] (ab280198) at 1/1000 dilution
Lane 1 : His-tagged human HDAC1 recombinant protein (aa1-482)
Lane 2 : His-tagged human HDAC2 recombinant protein (aa1-488)
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 55 kDa
Observed band size: 66 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
This antibody has no cross-reaction with human HDAC2.
These recombinant proteins were made in house. These two recombinant proteins were expressed from E.coli expression systems.
Exposure time: 10 seconds.
-
Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling HDAC1 with ab280198 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR23847-170] (ab280198)
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HDAC1 with ab280198 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human liver (PMID:18264140). The section was incubated with ab280198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
HDAC1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab280198 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280198 at 1/1000 dilution. 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: ab280198 IP in HeLa whole cell lysate 10 ug
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab280198 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
-
All lanes : Anti-HDAC1 antibody [EPR23847-170] (ab280198) at 1/5000 dilution
Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 55 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID:24551070).
Exposure time: 37 seconds.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR23847-170] (ab280198)
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling HDAC1 with ab280198 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse liver. The section was incubated with ab280198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR23847-170] (ab280198)
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling HDAC1 with ab280198 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat liver. The section was incubated with ab280198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.