Recombinant Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24219-30] to KDM3B / JMJD1B
- Suitable for: IHC-P, WB, Flow Cyt (Intra), ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-KDM3B / JMJD1B antibody [EPR24219-30]
See all KDM3B / JMJD1B primary antibodies -
Description
Rabbit monoclonal [EPR24219-30] to KDM3B / JMJD1B -
Host species
Rabbit -
Specificity
This antibody does not react with Human species for IHC.
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Tested applications
Suitable for: IHC-P, WB, Flow Cyt (Intra), ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HEK-293T, NIH/3T3 and PC-12 whole cell lysates; Mouse placenta and testis tissue lysates. IHC-P: Mouse testis tissue: Rat testis tissue. ICC/IF: HeLa and NIG/3T3 cells. Flow cyt: HeLa and NIH/3T3 cells.
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General notes
ab271046 does not react with Human species for IHC application.
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
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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
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24219-30 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab271046 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 |
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IHC-P |
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This antibody does not react with Human species for IHC. |
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WB |
1/1000. Detects a band of approximately 220 kDa (predicted molecular weight: 191 kDa).
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Flow Cyt (Intra) |
1/500.
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ICC/IF |
1/500.
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Notes |
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IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. This antibody does not react with Human species for IHC. |
WB
1/1000. Detects a band of approximately 220 kDa (predicted molecular weight: 191 kDa). |
Flow Cyt (Intra)
1/500. |
ICC/IF
1/500. |
Target
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Function
Histone demethylase that specifically demethylates 'Lys-9' of histone H3, thereby playing a central role in histone code. Demethylation of Lys residue generates formaldehyde and succinate. May have tumor suppressor activity. -
Tissue specificity
Ubiquitous. Highly expressed in placenta, skeletal muscle, kidney, heart and liver. -
Sequence similarities
Belongs to the JHDM2 histone demethylase family.
Contains 1 JmjC domain. -
Domain
Leu-Xaa-Xaa-Leu-Leu (LXXLL) motifs are known to mediate the association with nuclear receptors. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 51780 Human
- Entrez Gene: 277250 Mouse
- Entrez Gene: 682469 Rat
- Omim: 609373 Human
- SwissProt: Q7LBC6 Human
- SwissProt: Q6ZPY7 Mouse
- Unigene: 483486 Human
- Unigene: 479894 Mouse
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Alternative names
- 5qNCA antibody
- C5orf7 antibody
- JHDM2B antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)
Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling KDM3B / JMJD1B with ab271046 at 1/5000 dilution followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on rat testis. The section was incubated with ab271046 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 LeicaDS9800 (Bond™, Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labeling KDM3B / JMJD1B with ab271046 at 1/500 dilution, followed by 150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear 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.
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Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling KDM3B / JMJD1B with ab271046 at 1/500 dilution (0.1µg) (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, ab150081) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046) at 1/1000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate
Lane 2 : 293T ( human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 191 kDa
Observed band size: 220 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Lysates were made freshly and used in WB immediately to minimize protein degradation.
Exposure time: 3 minutes
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Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling KDM3B / JMJD1B with ab271046 at 1/500 dilution (0.1µg) (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, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling KDM3B / JMJD1B with ab271046 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubµue 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling KDM3B / JMJD1B with ab271046 at 1/5000 dilution followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on mouse testis (PMID: 26681924). The section was incubated with ab271046 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 LeicaDS9800 (Bond™, Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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All lanes : Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046) at 1/1000 dilution
Lane 1 : Mouse testis tissue lysate
Lane 2 : Mouse placenta tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 191 kDa
Observed band size: 220 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Lysates were made freshly and used in WB immediately to minimize protein degradation.
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 3 minutes.
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
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab271046 has not yet been referenced specifically in any publications.