Recombinant Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] - BSA and Azide free (ab251183)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18651] to KDM5A / Jarid1A / RBBP2 - BSA and Azide free
- Suitable for: WB, IP, Flow Cyt (Intra), ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
-
Product name
Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] - BSA and Azide free
See all KDM5A / Jarid1A / RBBP2 primary antibodies -
Description
Rabbit monoclonal [EPR18651] to KDM5A / Jarid1A / RBBP2 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, Flow Cyt (Intra), ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
General notes
ab251183 is the carrier-free version of ab194286.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18651 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab251183 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
WB |
Use at an assay dependent concentration. Detects a band of approximately 192 kDa (predicted molecular weight: 192 kDa).
|
|
IP |
Use at an assay dependent concentration.
|
|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
|
|
ICC/IF |
Use at an assay dependent concentration.
|
Notes |
---|
WB
Use at an assay dependent concentration. Detects a band of approximately 192 kDa (predicted molecular weight: 192 kDa). |
IP
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Target
-
Function
Histone demethylase that specifically demethylates 'Lys-4' of histone H3, thereby playing a central role in histone code. Does not demethylate histone H3 'Lys-9', H3 'Lys-27', H3 'Lys-36', H3 'Lys-79' or H4 'Lys-20'. Demethylates trimethylated and dimethylated but not monomethylated H3 'Lys-4'. May stimulate transcription mediated by nuclear receptors. May be involved in transcriptional regulation of Hox proteins during cell differentiation. May participate in transcriptional repression of cytokines such as CXCL12. -
Sequence similarities
Belongs to the JARID1 histone demethylase family.
Contains 1 ARID domain.
Contains 1 JmjC domain.
Contains 1 JmjN domain.
Contains 3 PHD-type zinc fingers. -
Domain
The GSGFP motif is required for the interaction with SUZ12. -
Cellular localization
Nucleus > nucleolus. Occupies promoters of genes involved in RNA metabolism and mitochondrial function. - Information by UniProt
-
Database links
- Entrez Gene: 5927 Human
- Entrez Gene: 214899 Mouse
- Omim: 180202 Human
- SwissProt: P29375 Human
- SwissProt: Q3UXZ9 Mouse
- Unigene: 76272 Human
- Unigene: 404761 Mouse
- Unigene: 463658 Mouse
-
Alternative names
- Histone demethylase JARID1A antibody
- JARID1A antibody
- Jumonji/ARID domain containing protein 1A antibody
see all
Images
-
This data was developed using ab194286, the same antibody clone in a different buffer formulation.
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: KDM5A knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab194286 observed at 240 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab194286 was shown to specifically recognize KDM5A in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when KDM5A knockout samples were examined. Wild-type and KDM5A knockout samples were subjected to SDS-PAGE. ab194286 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/2500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
-
This data was developed using ab194286, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling KDM5A / Jarid1A / RBBP2 with ab194286 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).The negative controls are as follows:
-ve control 1: ab194286 at 1/2000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution. -
All lanes : Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286) at 1/5000 dilution
Lane 1 : LLC (Mouse lung carcinoma) whole cell lysate
Lane 2 : Mouse testis lysate
Lane 3 : HEK-293 (Human epithelial cells from embryonic kidney) 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
Predicted band size: 192 kDa
Observed band size: 192 kDa
Exposure time: 30 secondsThis data was developed using ab194286, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
-
Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286) at 1/5000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 192 kDa
Observed band size: 192 kDa
Exposure time: 10 secondsThis data was developed using ab194286, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
-
Anti-KDM5A / Jarid1A / RBBP2 antibody [EPR18651] (ab194286) at 1/1000 dilution + NIH/3T3 (Mouse embryonic fibroblast cells) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 192 kDa
Observed band size: 192 kDa
Exposure time: 10 secondsThis data was developed using ab194286, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
-
This data was developed using ab194286, the same antibody clone in a different buffer formulation.KDM5A / Jarid1A / RBBP2 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab194286 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab194286 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution. Lane 1: HeLa whole cell lysate 10ug (Input). Lane 2: ab194286 IP in HeLa whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab194286 in HeLa whole cell lysate. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 5 seconds.
-
This data was developed using ab194286, the same antibody clone in a different buffer formulation.
Intracellular Flow Cytometry analysis of NIH/3T3 (mouse embryo) cells labelling KDM5A / Jarid1A / RBBP2 (red) with purified ab194286 at dilution of 1/1500. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody used was Rabbit Monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
Datasheet download
Certificate of Compliance
References (0)
ab251183 has not yet been referenced specifically in any publications.