Antibodies, Proteins, Kits and Reagents for Life Science

Anti-MBD3 antibody [EPR18258] (ab188401)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18258] to MBD3
Suitable for: WB, ICC/IF, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-MBD3 antibody [EPR18258]
See all MBD3 primary antibodies
Description

Rabbit monoclonal [EPR18258] to MBD3
Host species

Rabbit
Specificity

This antibody detects a doublet by western blot, which we believe corresponds to isoforms 1 and 2 of MBD3.

This product is only recommended for human IHC but not for mouse or rat.

Tested Applications & Species

Application	Species
ICC/IF	Mouse Human
IHC-P	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa, 293, F9, WEHI-3, T-47D, HepG2, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal kidney lysate; Mouse spleen lysate. ICC/IF: HeLa and NIH/3T3 cells.
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
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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18258
Isotype

IgG
Research areas

Western blot - Anti-MBD3 antibody [EPR18258] (ab188401)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: MBD3 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: F9 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab188401 observed at 33 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab188401 was shown to specifically react with MBD3 when MBD3 knockout samples were used. Wild-type and MBD3 knockout samples were subjected to SDS-PAGE. ab188401 and ab8245 (loading control to GAPDH) were diluted 1/5000 and 1/10000 and incubated respectively overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD3 antibody [EPR18258] (ab188401)

Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling MBD3 with ab188401 at 1/200 dilution (6.17 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the human stomach. The section was incubated with ab188401 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Immunocytochemistry/ Immunofluorescence - Anti-MBD3 antibody [EPR18258] (ab188401)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling MBD3 with ab188401 at 1/500 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 ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab188401 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Western blot - Anti-MBD3 antibody [EPR18258] (ab188401)

All lanes : Anti-MBD3 antibody [EPR18258] (ab188401) at 1/5000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : 293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 3 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
Lane 4 : WEHI-3 (Mouse leukemia cell line) whole cell lysate
Lane 5 : T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate
Lane 6 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 33, 29 kDa
Observed band size: 29,33 kDa
why is the actual band size different from the predicted?

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.
The 33 and 29 kDa bands correspond to Isoforms 1 and 2.
Immunocytochemistry/ Immunofluorescence - Anti-MBD3 antibody [EPR18258] (ab188401)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling MBD3 with ab188401 at 1/500 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 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab188401 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD3 antibody [EPR18258] (ab188401)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling MBD3 with ab188401 at 1/200 dilution (6.17 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the human pancreas (PMID: 27898661). The section was incubated with ab188401 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Western blot - Anti-MBD3 antibody [EPR18258] (ab188401)

All lanes : Anti-MBD3 antibody [EPR18258] (ab188401) at 1/2000 dilution

Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 33, 29 kDa
Observed band size: 29,33 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
The 33 and 29 kDa bands correspond to Isoforms 1 and 2.
Western blot - Anti-MBD3 antibody [EPR18258] (ab188401)

Anti-MBD3 antibody [EPR18258] (ab188401) at 1/2000 dilution + Human fetal kidney lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 33, 29 kDa
Observed band size: 29,33 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-MBD3 antibody [EPR18258] (ab188401)

Anti-MBD3 antibody [EPR18258] (ab188401) at 1/2000 dilution + Mouse spleen lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 33, 29 kDa
Observed band size: 29,33 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
The 33 and 29 kDa bands correspond to Isoforms 1 and 2.
Anti-MBD3 antibody [EPR18258] (ab188401)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MBD3 antibody [EPR18258] (ab188401)

Key features and details

Overview

Product name

Description

Host species

Specificity

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Images

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