Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µl 298 185 ₸

Order now and get it on Tuesday March 02, 2021

Anti-CD3D antibody [EPR20544] (ab213362)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20544] to CD3D
Suitable for: WB, IHC-P, IHC-Fr, Flow Cyt, IP
Reacts with: Mouse

Product name

Anti-CD3D antibody [EPR20544]
See all CD3D primary antibodies
Description

Rabbit monoclonal [EPR20544] to CD3D
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
IHC-Fr	Mouse
IHC-P	Mouse
IP	Mouse
WB	Mouse

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Mouse thymus and lymph node lysates. IHC-P: Mouse spleen and thymus tissues. IHC-Fr: Mouse spleen tissue. Flow Cyt: Mouse spleen cells. IP: Mouse thymus lysate.
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, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20544
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EPR20544] (ab213362)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD3D with ab213362 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Cytoplasmic staining on T cells of mouse splenic periarterial lymphatic sheath 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) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Frozen sections) - Anti-CD3D antibody [EPR20544] (ab213362)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen tissue labeling CD3D with ab213362 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Cytoplasmic and membranous staining on mouse splenic periarterial lymphatic sheath is observed.

The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Western blot - Anti-CD3D antibody [EPR20544] (ab213362)

All lanes : Anti-CD3D antibody [EPR20544] (ab213362) at 1/1000 dilution

Lane 1 : Mouse thymus lysate
Lane 2 : Mouse lymph node 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: 19 kDa
Observed band size: 24 kDa
why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 15 seconds; Lane 2: 3 minutes.
Flow Cytometry - Anti-CD3D antibody [EPR20544] (ab213362)

Flow cytometric analysis of mouse spleen cells labeling CD3D with ab213362 at 1/600 dilution (right panel), compared with a rabbit monoclonal IgG isotype control (ab172730) (left panel). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

Firstly, cells surface was stained with anti-mouse CD3 Alexa Fluor 647 (Y axis), then cells were fixed with 4% PFA followed by intracellular staining with ab213362. The same population of cells were stained by both: anti-mouse CD3 and anti-mouse CD3D (ab213362) antibodies.
Immunoprecipitation - Anti-CD3D antibody [EPR20544] (ab213362)

CD3D was immunoprecipitated from 0.35 mg of mouse thymus lysate with ab213362 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab213362 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1: Mouse thymus lysate, 10 µg (Input).

Lane 2: ab213362 IP in mouse thymus lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213362 in mouse thymus lysate .

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EPR20544] (ab213362)

Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling CD3D with ab213362 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Cytoplasmic and membranous staining on T cells of mouse thymus medulla 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) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-CD3D antibody [EPR20544] (ab213362)

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