Recombinant Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18667] to EHMT2/G9A + EHMT1/GLP
- Suitable for: IHC-P, WB, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667]
See all EHMT2/G9A+EHMT1/GLP primary antibodies -
Description
Rabbit monoclonal [EPR18667] to EHMT2/G9A + EHMT1/GLP -
Host species
Rabbit -
Specificity
Not suitable for IP-Human and IP-Mouse.
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Tested applications
Suitable for: IHC-P, WB, ICC/IF, Flow Cyt (Intra)more details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human fetal kidney lysate; HEK-293, HeLa, K562, MCF7 and PC-3 whole cell lysates; EHMT2 recombinant protein, Wild-type HAP1 cell lysate, HEK-293T; NIH/3T3, PC-12 and Raw 264.7 cell lysates. Flow Cyt (Intra): HeLa cells. IHC-P: Human tonsil tissue; ICC/IF: HeLa and NIH/3T3 cells.
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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
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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: 7.2
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
EPR18667 -
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 ab194299 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 |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Detects a band of approximately 170 kDa (predicted molecular weight: 141, 132 kDa).
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ICC/IF |
1/1000.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 170 kDa (predicted molecular weight: 141, 132 kDa). |
ICC/IF
1/1000. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
Target
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Cellular localization
EHMT2/G9A: Nucleus. Chromosome. Associates with euchromatic regions. Does not associate with heterochromatin. EHMT1/GLP: Nucleus. Chromosome. Associates with euchromatic regions. -
Database links
- Entrez Gene: 10919 Human
- Entrez Gene: 79813 Human
- Entrez Gene: 110147 Mouse
- Entrez Gene: 77683 Mouse
- Entrez Gene: 361798 Rat
- Entrez Gene: 362078 Rat
- Omim: 604599 Human
- Omim: 607001 Human
see all
Images
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All lanes : Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate
Lane 4 : Raw 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) 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: 141, 132 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
Exposure Time: Lane 1: 15 seconds, Lane 2-4: 37 seconds.
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
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All lanes : Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : EHMT1 knockout HAP1 cell lysate
Lane 3 : Wild-type HAP1 Nuclear cell lysate
Lane 4 : HeLa cell lysate
Lane 5 : HeLa Nuclear cell lysate
Lane 6 : Wild-type HEK-293T cell lysate
Lane 7 : Wild-type HEK-293T nuclear cell lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preabsorbed (ab216776) at 1/20000 dilution
Predicted band size: 141, 132 kDa
Observed band size: 150-170 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab194299 was shown to bind specifically to EHMT1/GLP and EHMT2/G9A. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299) at 1/1000 dilution + Human fetal kidney lysate at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution
Developed using the ECL technique.
Predicted band size: 141, 132 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling EHMT2/G9A + EHMT1/GLP (red) with purifiedab194299at a dilution of 1/150. Goat anti rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/2000. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.
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All lanes : Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299) at 1/5000 dilution
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 141, 132 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lanes 1 & 2: 20 seconds; Lane 3: 30 seconds.
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Lane 1 : Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299) at 1/5000 dilution
Lane 2 : Anti His-Tag®.
All lanes : EHMT2 recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 141, 132 kDa
Observed band size: 30,60 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
EHMT2 recombinant protein fragment with His-Tag®.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling EHMT2/G9A + EHMT1/GLP with ab194299 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human tonsil is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling EHMT2/G9A + EHMT1/GLP with ab194299 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (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: ab194299 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution. -
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling EHMT1/GLP + EHMT2/G9A with ab194299 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3. 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: ab194299 at 1/1000 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab194299 has not yet been referenced specifically in any publications.