Synaptic Systems

Cat. No.: 555 003

Amount: 50 µg

Price: $380.00

Please note that import duties may still apply, which will be billed to you directly by the shipping carrier.

CD28 staining of a human placenta

WB
(1 img)

IHC-P (FFPE)
(4 img)

Background

CD28 is a central co-stimulatory receptor required for optimal T-cell activation, expansion, and survival. Constitutively expressed on most naïve T-cells, CD28 binds the B7 family ligands CD80 (B7-1) and CD86 (B7-2) on antigen-presenting cells, delivering the “second signal” necessary to complement T-cell receptor (TCR) recognition of antigen and prevent anergy (1, 2). Upon ligation, CD28 amplifies intracellular signaling pathways including phosphoinositide 3-kinase (PI3K), Akt, NF-κB, and MAPK, thereby enhancing interleukin-2 (IL-2) production, cell-cycle progression, metabolic fitness, and resistance to apoptosis (3, 4). In addition to its role in normal immune responses, dysregulated CD28 signaling contributes to autoimmunity, chronic inflammation, and transplant rejection, while its manipulation has become highly relevant in cancer immunotherapy and immune modulation (5). The importance of CD28 is also underscored in chimeric antigen receptor (CAR) T-cell design, where incorporation of the CD28 intracellular signaling domain promotes robust effector activity and functional persistence (6, 7).

1 ) Linsley PS et al., J Exp Med (1991)
2 ) Harding FA et al., Nature (1992)
3 ) Fraser JD et al., Science (1991)
4 ) Boomer JS et al., Cold Spring Harb Perspect Biol (2010)
5 ) Esensten JH et al., Immunity (2016)
6 ) Maher J et al., Nat Biotechnol (2002)
7 ) Sadelain M et al., Cancer Discov (2013)

Protocols

WB

IHC-P (FFPE)

Cat. No. 555 003	50 µg specific antibody, lyophilized. Affinity purified with the immunogen. Albumin and azide were added for stabilization. For reconstitution add 50 µl H₂O to get a 1mg/ml solution in PBS. Then aliquot and store at -20°C to -80°C until use. Antibodies should be stored at +4°C when still lyophilized. Do not freeze!
Applications	WB: 1 : 500 (fluorescent detection) gallery IP: not tested yet ICC: not tested yet IHC: not tested yet IHC-P (FFPE): yes (see remarks) gallery Western blot (WB); separation of proteins by PAGE and subsequent transfer to a membrane. Detection of target molecules is carried out with antibodies. Some antibodies require special sample preparation steps. For details, please refer to the “Remarks” section. Immunoprecipitation (IP); Immunoisolation or pulldown of a target molecule using an antibody. For details and product specific hints, please refer to the ”Remarks” section. Immunocytochemistry (ICC) on 4% PFA fixed cells. Immunoreactivity is usually revealed by fluorescence. Some antibodies require special fixation methods. For details, please refer to the “Remarks” section. Immunohistochemistry (IHC) on 4% PFA perfusion fixed tissue with 24h PFA post fixation. Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate. Some antibodies require special fixation methods or antigen retrieval steps. For details, please refer to the ”Remarks” section. Immunohistochemistry (IHC-P) of formalin fixed, paraffin embedded (FFPE) tissue (some antibodies require special antigen retrieval steps, please refer to the ”Remarks” section). Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate.
Immunogen	Synthetic peptide corresponding to residues near the carboxy terminus of human CD28 (UniProt Id: P10747)
Reactivity	Reacts with: human (P10747). Other species not tested yet.
Remarks	IHC-P (FFPE): Antigen retrieval with Tris-EDTA buffer pH 9 is recommended for chromogenic detection. For chromogenic detection, an optimized AGR time of 20 minutes is recommended for best results.
Data sheet	Data Sheet 555_003 Safety Data Sheet (SDS)

CD28 antibody - 555 003