Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)
![Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)](https://www.abcam.com/ps/products/254/ab254009/Images/ab254009-389741-anti-dnmt3a-antibody-epr18455-94-western-blot-human-lysate.jpg "Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18455-94] to Dnmt3a - BSA and Azide free
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free
See all Dnmt3a primary antibodies -
Description
Rabbit monoclonal [EPR18455-94] to Dnmt3a - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: ChIP,Flow Cyt,ICC/IF or 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: HAP1, HEK-293T, HeLa and NIH/3T3 whole cell lysates. IHC-P: Human breast carcinoma and placenta tissue; mouse colon tissue; rat colon tissue.
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General notes
Ab254009 is the carrier-free version of ab227823. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab254009 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
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
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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
EPR18455-94 -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)All lanes : Anti-Dnmt3a antibody [EPR18455-94] (ab227823) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : Dnmt3a knockout HeLa cell lysate
Lane 3 : HEK-293 cell lysate
Lane 4 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 102 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab227823).
Lanes 1-4: Merged signal (red and green). Green - ab227823 observed at 125 kDa. Red - loading control ab8245 observed at 37 kDa.
ab227823 Anti-Dnmt3a antibody [EPR18455-94] was shown to specifically react with Dnmt3a in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261793 (knockout cell lysate ab257128) was used. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab227823 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)](https://www.abcam.com/ps/products/254/ab254009/Images/ab254009-343882-Anti-Dnmt3a-antibody-western-blot-hap1-human.jpg)
Western blot - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)All lanes : Anti-Dnmt3a antibody [EPR18455-94] (ab227823) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate, 20 ug
Lane 2 : Dnmt3a knockout HAP1 whole cell lysate, 20 ug
Lane 3 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate, 20 ug
Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lane 5 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate, 20 ug
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 102 kDaBlocking and dilution buffer: 5% NFDM/TBST.
ab227823 was shown to specifically react with wild-type HAP1 cells as signal was lost in Dnmt3a knockout cells. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab227823 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
The observed bands ranging from 75~125kDa represent different Dnmt3a isoforms.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227823).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)](https://www.abcam.com/ps/products/254/ab254009/Images/ab254009-4-227823IHC4.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the rat colon (PMID: 24837369) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227823).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.
The section was incubated with ab227823 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)](https://www.abcam.com/ps/products/254/ab254009/Images/ab254009-3-227823IHC3.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the mouse colon (PMID: 24837369) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227823).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.
The section was incubated with ab227823 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)](https://www.abcam.com/ps/products/254/ab254009/Images/ab254009-2-227823IHC2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the human placenta (PMID: 22413869) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227823).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.
The section was incubated with ab227823 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)](https://www.abcam.com/ps/products/254/ab254009/Images/ab254009-1-227823IHC1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling Dnmt3a with ab227823 at 1/200 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the human breast carcinoma (PMID: 24464625) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227823).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.
The section was incubated with ab227823 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
-
![Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)](https://www.abcam.com/ps/products/254/ab254009/Images/ab254009-7-benefits-of-recombinant-antibodies.png)
Anti-Dnmt3a antibody [EPR18455-94] - BSA and Azide free (ab254009)