Recombinant Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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

Carrier Free

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.
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.
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.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR24219-30
Isotype

IgG
Research areas

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
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.

Notes
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.

Application notes

Is unsuitable for IP.

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.
Target information above from: UniProt accession Q7LBC6 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
Alternative names
- 5qNCA antibody
- C5orf7 antibody
- JHDM2B antibody
- JmjC domain containing histone demethylation protein 2B antibody
- JmjC domain-containing histone demethylation protein 2B antibody
- jmjd1b antibody
- Jumonji domain containing 1B antibody
- Jumonji domain containing protein 1B antibody
- Jumonji domain-containing protein 1B antibody
- KDM3B antibody
- KDM3B_HUMAN antibody
- KIAA1082 antibody
- Lysine (K) specific demethylase 3B antibody
- Lysine-specific demethylase 3B antibody
- NET22 antibody
- Nuclear protein 5qNCA antibody
see all

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.
Immunocytochemistry/ Immunofluorescence - Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)

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 Fluo^r® 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.
Flow Cytometry (Intracellular) - Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)

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.
Western blot - Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)

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
Flow Cytometry (Intracellular) - Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)

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.
Immunocytochemistry/ Immunofluorescence - Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)

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.
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.
Western blot - Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)

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.
Anti-KDM3B / JMJD1B antibody [EPR24219-30] (ab271046)

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.

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab271046? Please let us know so that we can cite the reference in this datasheet.

ab271046 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Compatible Secondaries

Isotype control

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As