JMJD3 Antibody

Contact us
ProSci
Product Code: PSI-6657
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-6657-0.02mg0.02mg£150.00
Quantity:
PSI-6657-0.1mg0.1mg£449.00
Quantity:
Prices exclude any Taxes / VAT

Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunofluorescence (IF)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

1 / 8
<strong>Figure 1 Western Blot Validation in Human, Mouse and Rat Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: JMJD3, 6657, (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
2 / 8
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines </strong><br> Loading: 15 μg of lysates per lane. Antibodies: JMJD3 6657 (1 μg/mL), JMJD3 14-618, (2 μg/mL) and beta-actin 3779 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
3 / 8
<strong>Figure 3 Western Blot Validation in HeLa Cell Lysate</strong><br> Loading: 15 μg of lysates per lane. Antibodies: JMJD3, 6657 (0.25 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
4 / 8
<strong>Figure 4 Western Blot Validation in K562 Cell Lysate </strong><br> Loading: 15 μg of lysates per lane. Antibodies: : JMJD3, 6657 (0.5 μg/mL) , 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: in the absence and Lane 2: in the presence of blocking peptide
5 / 8
<strong>Figure 5 Western Blot Validation in Mouse A-20 Cell Lysate </strong><br> Loading: 15 μg of lysates per lane. Antibodies: : JMJD3, 6657 (0.5 μg/mL) , 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: in the absence and Lane 2: in the presence of blocking peptide
6 / 8
<strong>Figure 6 Western Blot Validation in HepG2 Cell Lysate </strong><br> Loading: 15 μg of lysates per lane. Antibodies: : JMJD3, 6657, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 0.25 μg/mL Lane 2: 0.5 μg/mL Lane 3: 0.25 μg/mL with blocking peptide
7 / 8
<strong>Figure 7 Immunocytochemistry Validation of JMJD3 in K562 Cells</strong><br> Immunocytochemical analysis of K562 cells using anti-JMJD3 antibody (6657) at 2.5 μg/ml. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
8 / 8
<strong>Figure 8 Immunofluorescence Validation of JMJD3 in K562 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling JMJD3 with 6657 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
❮❯

<strong>Figure 1 Western Blot Validation in Human, Mouse and Rat Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: JMJD3, 6657, (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines </strong><br> Loading: 15 μg of lysates per lane. Antibodies: JMJD3 6657 (1 μg/mL), JMJD3 14-618, (2 μg/mL) and beta-actin 3779 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Western Blot Validation in HeLa Cell Lysate</strong><br> Loading: 15 μg of lysates per lane. Antibodies: JMJD3, 6657 (0.25 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Western Blot Validation in K562 Cell Lysate </strong><br> Loading: 15 μg of lysates per lane. Antibodies: : JMJD3, 6657 (0.5 μg/mL) , 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: in the absence and Lane 2: in the presence of blocking peptide
<strong>Figure 5 Western Blot Validation in Mouse A-20 Cell Lysate </strong><br> Loading: 15 μg of lysates per lane. Antibodies: : JMJD3, 6657 (0.5 μg/mL) , 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: in the absence and Lane 2: in the presence of blocking peptide
<strong>Figure 6 Western Blot Validation in HepG2 Cell Lysate </strong><br> Loading: 15 μg of lysates per lane. Antibodies: : JMJD3, 6657, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 0.25 μg/mL Lane 2: 0.5 μg/mL Lane 3: 0.25 μg/mL with blocking peptide
<strong>Figure 7 Immunocytochemistry Validation of JMJD3 in K562 Cells</strong><br> Immunocytochemical analysis of K562 cells using anti-JMJD3 antibody (6657) at 2.5 μg/ml. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 8 Immunofluorescence Validation of JMJD3 in K562 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling JMJD3 with 6657 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Documents

Further Information

Additional Names:
JMJD3 Antibody: JMJD3, JMJD3, KIAA0346, Lysine-specific demethylase 6B, JmjC domain-containing protein 3
Application Note:
WB: 0.25-4 μg/mL; ICC: 2.5 μg/mL; IF: 20 μg/mL.

Antibody validated: Western Blot in human, mouse and rat samples; Immunocytochemistry in human samples; Immunofluorescence in human samples. All other applications and species not yet tested.
Background:
JMJD3 Antibody: The Jumonji domain-containing protein 3 (JMJD3) functions as a trimethylation-specific demethylase, converting the trimethylated histone H3 Lys27 residue to the dimethylated form, and is thought to also function as a transcriptional repressor. JMJD3 plays a central role in regulation of posterior development, by regulating HOX gene expression. It is involved in inflammatory response by participating in macrophage differentiation in case of inflammation by regulating gene expression and macrophage differentiation. JMJD3 can also interact with and demethylate p53, resulting in its stabilization and localization to the nucleus in mouse embryo fibroblasts during neural stem cell differentiation.
Background References:
  • Agger et al. Nature 2007; 449:731-4.
  • De Santa et al. Cell 2007; 130:1083-94.
  • Sola et al. PLoS One 2011; 6:e18421.
Buffer:
JMJD3 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Immunogen:
Anti-JMJD3 antibody (6657) was raised against a peptide corresponding to 16 amino acids near the carboxy terminus of human JMJD3.

The immunogen is located within amino acids 1370-1420 of JMJD3.
ISOFORMS:
Human JMJD3 has 2 isoforms, including isoform 1 (1682aa, 181kD) and isoform 2 (1643aa, 177kD). Mouse JMJD3 has one isoform (1641aa, 176kD) and rat JMJD3 also has one isoform (1356aa, 146kD). 6657 can detect human, mouse and rat.
NCBI Gene ID #:
23135
NCBI Official Name:
lysine (K)-specific demethylase 6B
NCBI Official Symbol:
KDM6B
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 180

Observed: 180 kD
Protein Accession #:
EAW90126
Protein GI Number:
122937251
Purification:
JMJD3 Antibody is affinity chromatography purified via peptide column.
Research Area:
Stem Cell
SPECIFICITY:
This antibody is specific for JMJD3 and will not recognize other JMJD proteins.
Swissprot #:
O15054
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

Independent Antibody Validation in Cell lines (Figure 2) shows similar JMJD3 expression profile in human, mouse and rat cell lines detected by two independent anti-IFN-beta antibodies that recognize different epitopes, 6657 against C-terminus domain and 14-618 against N-terminus domain.  IFN-beta proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. 

Related Products

Product NameProduct CodeSupplier 
JMJD3 PeptidePSI-6657PProSci Summary Details