CD3 Rabbit Monoclonal Antibody(ARA817)

Datasheet

MSDS

Key features and details

Target: CD3
Source/Host: Rabbit
Reactivity: Human
Clonality: Monoclonal
Applications: WB,IHC,IF/ICC,FC,IP
Conjugation: Unconjugated
Storage: at-20°C

Brand:

CAT.NO. : ARA6607

US$ Please choose

Size:

100μL

Trail, Bulk size or Custom requests Please contact us

Product Details

Background

Part of the TCR-CD3 complex present on T-lymphocyte cell surface that plays an essential role in adaptive immune response. When antigen presenting cells (APCs) activate T-cell receptor (TCR), TCR-mediated signals are transmitted across the cell membrane by the CD3 chains CD3D, CD3E, CD3G and CD247/CD3Z. All CD3 chains contain immunoreceptor tyrosine-based activation motifs (ITAMs) in their cytoplasmic domain. Upon TCR engagement, these motifs become phosphorylated by Src family protein tyrosine kinases LCK and FYN, resulting in the activation of downstream signaling pathways activate T-cell receptor (TCR), TCR-mediated signals are transmitted across the cell membrane by the CD3 chains CD3D, CD3E, CD3G and CD247/CD3Z.

Application

To ensure optimal assay performance, AREX recommends conducting reagent titration tailored to each testing system for optimal detection results.

WB	1:10000-1:50000
IHC	1:100-1:200
IF/ICC	1:50-1:2000
FC	1:200-1:1000
IP	1:15

*Results are sample-specific. Please refer to your local assay conditions and test parameters for reference.

Overview

Description	Rabbit Monoclonal Antibody to CD3
Antibody Type	Primary antibody
Predicted MW	23kDa
Immunogen	A synthetic peptide corresponding to residues on the N-terminus of human CD3 was used as an immunogen.
Purification	ProA affinity purified IgG
Form/Buffer	PBS 59%, Sodium azide 0.01%, Glycerol 40%, BSA 0.92%.
Alternative Names	T3E; T-cell surface glycoprotein CD3 epsilon chain; T-cell surface antigen T3/Leu-4 epsilon chain; CD3e
Gene Symbol	CD3E
Entrez Gene	916(Human)
Swissprot	P07766

*AREX continuously optimizes our products. Webpage content may not reflect the latest updates. For inquiries, please contact info@arexbio.com or your local distributor.
*Clone Number, Reactivity, Source/Host and Clonality can be found in the product name and Key Features section above.

Data

All lane: Anti-CD3 antibody at 1:10,000 dilution
Predicted MW: 23 kDa
Observed MW: 23 kDa
Lane 1: JurKat
Lysate at 10 µg per lane
2nd Ab:
G&R HRP(H+L) 1:4,000
Exposure: 60s

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD3 with ARA817 at 1:200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0.

ARA817 staining CD3 in Jurkat cells by IF/ICC (immunofluorescence/immunocytochemistry). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% goat serum for half an hour at room temperature. Samples were incubated with primary antibody (1:50) at 4°C. An Alexa Fluor® 488-conjugated Goat Anti-Rabbit IgG polyclonal was used as the secondary antibody (1:500). DAPI (blue) was used as the nuclear counter stain.
Control: PBS and secondary antibody, An Alexa Fluor® 488-conjugated Goat Anti-Rabbit IgG (1:500).

Overlay histogram showing Jurkat cells stained with ARA817 (Blue). The cells were fixed with 4% paraformaldehyde for 10 min. The cells were then incubated in the antibody (ARA817, 1:1,000 dilution) in 1x PBS/1% BSA for 30 min at room temperature. The secondary antibody used was a Goat Anti-Rabbit Alexa Fluor® 488 (IgG H+L) at 1:2,000 dilution for 20 min at room temperature. Unlabelled sample (Red) was used as a control.

CD3 was immunoprecipitated from 0.4mg of Molt-4 whole cell lysate with ARA817 at 1:15 dilution.
2nd Ab:
GAR HRP for IP 1:500
Lane 1: ARA817 IP in Molt-4 whole cell lysate
Lane 2: Rabbit IgG instead of ARA817 in Molt-4 whole cell lysate
Lane 3: Molt-4 whole cell lysate, 10 µg (input)
Exposure: 120s

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Note

For Research Use Only. Not for diagnostic, therapeutics, prophylactic or in vivo use.

FAQs

What are the main types of research antibodies and how do they differ?

Research antibodies are mainly divided into monoclonal antibodies and polyclonal antibodies. Monoclonal antibodies typically offer higher specificity and better batch-to-batch consistency, while polyclonal antibodies often provide stronger affinity but may show more variation between batches. The choice depends on your specific experimental needs.

How can I tell if a research antibody is suitable for my experiment?

It is recommended to carefully review the product datasheet for validated applications, species reactivity, recommended dilutions, and published references. For new antibodies, performing a small-scale validation with positive control samples is usually helpful.

Can improper storage of research antibodies affect experimental results?

Yes. Antibodies are sensitive to temperature, repeated freeze-thaw cycles, and contamination. Improper storage may lead to reduced activity, increased background, or weaker signals. It is best to follow the storage instructions provided in the product datasheet.

Why doesn’t the recommended dilution in the datasheet work well in my experiment?

The recommended dilution is based on the supplier’s test conditions. Factors such as sample type, fixation method, and detection system in your lab can influence the optimal working concentration. Performing a dilution series optimization in your own system is often necessary.

What precautions should I take when using a newly purchased research antibody for the first time?

It is advisable to briefly centrifuge the antibody (especially concentrated or lyophilized ones), then perform a small-scale pilot experiment using the recommended conditions. Recording the batch number and usage date is also helpful for future tracking.

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