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Your Position: Home > Immune Checkpoint Proteins > CTLA-4 > Biotinylated Human CTLA-4 Protein, Fc,Avitag

Biotinylated Human CTLA-4 Protein, Fc,Avitag™

INTRODUCTION

QC & Assay Protocol

Citation & Background

Synonym
CTLA4,CD152
Source
Biotinylated Human CTLA-4, Fc,Avitag (CT4-H82F3) is expressed from human 293 cells (HEK293). It contains AA Ala 37 - Phe 162 (Accession # P16410-1).
Predicted N-terminus: Ala 37
Request for sequence
Molecular Characterization
This protein carries a human IgG1 Fc tag at the C-terminus, followed by a Avi tag (Avitag™).
The protein has a calculated MW of 41.8 kDa. The protein migrates as 48-60 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Biotinylation
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
Biotin:Protein Ratio
The biotin to protein ratio is 0.5-1 as determined by the HABA assay.

Endotoxin
Less than 1.0 EU per μg by the LAL method.
Purity
>95% as determined by reduced SDS-PAGE.
Formulation
Lyophilized from 0.22 μm filtered solution in Tris with Glycine, Arginine and NaCl, pH7.5. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
No activity loss was observed after storage at:
1. -20^°C to -70^°C for 12 months in lyophilized state;
2. -70^°C for 3 months under sterile conditions after reconstitution.

SDS-PAGE

Biotinylated Human CTLA-4, Fc,Avitag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.

Bioactivity-ELISA

Biotinylated Human CTLA-4, Fc,AvitagBiotinylated Human CTLA-4, Fc,Avitag (Cat. No. CT4-H82F3) ELISA bioactivity

Immobilized Human B7-2, Fc Tag (Cat. No. CD6-H5257) at 5 μg/mL (100 μL/well) can bind Biotinylated Human CTLA-4, Fc,Avitag (Cat. No. CT4-H82F3) with a linear range of 0.6-10 ng/mL (QC tested).

Protocol

Immobilized Human B7-1, Fc Tag (Cat. No. B71-H5259) at 5 μg/mL (100 μL/well) can bind Biotinylated Human CTLA-4, Fc,Avitag (Cat. No. CT4-H82F3) with a linear range of 0.6-10 ng/mL (Routinely tested).

Protocol

Immobilized Cynomolgus / Rhesus macaque B7-2, His Tag (Cat. No. CD6-C52H5) at 5 μg/mL (100 μL/well) can bind Biotinylated Human CTLA-4, Fc,Avitag (Cat. No. CT4-H82F3) with a linear range of 0.02-0.625 μg/mL (Routinely tested).

Protocol

Immobilized Biotinylated Human CTLA-4, Fc,Avitag (Cat. No. CT4-H82F3) at 1 μg/mL (100 μL/well) on streptavidin precoated (0.5 μg/well) plate, can bind Ipilimumab with a linear range of 0.2-2 ng/mL (Routinely tested).

Protocol

Serial dilutions of Ipilimumab neutralizing antibody (1:2 serial dilutions, from 40 μg/mL to 0.019 μg/mL) were added into Human B7-1, Fc Tag (Cat. No. B71-H5259): Biotinylated Human CTLA-4, Fc,Avitag (Cat. No. CT4-H82F3) binding reactions. The assay was performed according to the above described protocol. Background was subtracted from data points before curve fitting.

Protocol

Serial dilutions of Ipilimumab neutralizing antibody (1:2 serial dilutions, from 40 μg/mL to 0.019 μg/mL) were added into Human B7-2, Fc Tag (Cat. No. CD6-H5257): Biotinylated Human CTLA-4, Fc,Avitag (Cat. No. CT4-H82F3) binding reactions. The assay was performed according to the above described protocol. Background was subtracted from data points before curve fitting.

Protocol

Citations
- Revealing the atomistic details behind the binding of B7–1 to CD28 and CTLA-4: A comprehensive protein-protein modelling study
  Authors: G Aravindhan, et al.
  Journal: BBA-GEN SUBJECTS 2018
  Application: ELISA
  Request for Full-text
Background
CTLA-4 (Cytotoxic T-Lymphocyte Antigen 4) is also known as CD152 (Cluster of differentiation 152), is a protein receptor that downregulates the immune system. CTLA4 is a member of the immunoglobulin superfamily, which is expressed on the surface of Helper T cells and transmits an inhibitory signal to T cells. The protein contains an extracellular V domain, a transmembrane domain, and a cytoplasmic tail. Alternate splice variants, encoding different isoforms. CTLA4 is similar to the T-cell co-stimulatory protein, CD28, and both molecules bind to CD80 and CD86, also called B7-1 and B7-2 respectively, on antigen-presenting cells. CTLA4 transmits an inhibitory signal to T cells, whereas CD28 transmits a stimulatory signal. Intracellular CTLA4 is also found in regulatory T cells and may be important to their function. Fusion proteins of CTLA4 and antibodies (CTLA4-Ig) have been used in clinical trials for rheumatoid arthritis.
References
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.

Order Details

CT4-H82F3-25ug

Price(USD) : $350.00

CT4-H82F3-200ug

Price(USD) : $1380.00

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English Name	Research Code	Research Phase	Company	First Brand Name	First Approved Country	First Indication	First Approved Company	First Approved Date	Indications	Clinical Trials
Ipilimumab	BMS-734016; MDX-010; MDX-101; 10D1; Anti-CTLA-4 Mab; Mab-10D14; MDX-CTLA-4,BMS734016,MDX010,MDX101	Approved	Bristol-Myers Squibb, Ono Pharmaceutical	YERVOY	fda	Metastatic melanoma	BRISTOL MYERS SQUIBB	2011-03-25	Renal cell carcinoma, Metastatic melanoma, Melanoma	Details

English Name	Research Code	Research Phase	Company	Indications	Clinical Trials
KN-044	KN-044,KN044; KN 044	Phase Ⅰ	Alphamab	Solid tumours	Details
ADU-1604	ADU-1604,ADU 1604,ADU1604	Phase Ⅰ	Aduro BioTech	Cancer	Details
Ipilimumab/IDD-1 combination vaccine (Medarex/IDM)		Phase Not Specified	Medarex, IDM Pharma	Prostate cancer	Details
Abatacept (Orban Biotech)		Phase Ⅱ	The National Institute of Diabetes and Digestive and Kidney Diseases, Orban Biotech	Type 1 diabetes	Details
BMS-986249	BMS-986249	Phase Ⅱ	CytomX Therapeutics, Bristol-Myers Squibb	Solid tumours	Details
MK-1308	MK-1308	Phase Ⅰ	Merck Sharp & Dohme	Solid tumours	Details
AK-104	AK-104,AK104	Phase Ⅱ	Akeso Biopharma	Solid tumours	Details
Anti-CTLA-4 monoclonal antibody (Regeneron)	REGN-4659	Phase Ⅰ	Regeneron Pharmaceutical	Non small cell lung cancer (NSCLC)	Details
ONC-392	ONC-392	Phase Ⅱ	OncoImmune	Non small cell lung cancer (NSCLC), Solid tumours	Details
KN-046	KN-046	Phase Ⅱ	Alphamab	Gastric cancer, Non small cell lung cancer (NSCLC), oesophageal cancer, Breast cancer, Solid tumours, Lymphoma	Details
HBM-4003	HBM-4003	Phase Ⅰ	Harbour BioMed	Solid tumours	Details
CS-1002	CS-1002,CS1002	Phase Ⅰ	CStone Pharmaceuticals	Solid tumours	Details
RG 2077		Phase Ⅰ	Repligen	Diabetes, Graft versus host disease, Systemic lupus erythematosus, Multiple sclerosis (MS), Rheumatoid arthritis (RA), Idiopathic thrombocytopenic purpura (ITP)	Details
PRS-010 (Pieris)	Pieries-110; PRS-010; PRS-030; PRS-055; PRS-056; PRS-070; PRS-090; PRS-110; PRS-130; PRS-190	Preclinical	Pieris Pharmaceuticals, Syngenta	Cancer	Details
Recombinant human anti-CTLA-4 monoclonal antibody (Mab-Venture/ShuangLu Pharmaceutical)		Phase Ⅰ	Mab-Venture, Beijing ShuangLu Pharmaceutical	Solid tumours	Details
XmAb-22841	XmAb-22841	Phase Ⅰ	Xencor	Cancer	Details
ATOR-1144	ATOR-1144	Preclinical	Alligator Bioscience	hematological malignancies, Solid tumours	Details
XmAb20717	XmAb-20717,XmAb 20717	Phase Ⅰ	Xencor	Solid tumours	Details
10B-10	10B-10	Preclinical	Crown Bioscience, Jiangsu Qyuns Biotech, CBT	Cancer	Details
Recombinant anti-CTLA-4 human monoclonal antibody (Hualan Biological Engineering)		Phase Ⅰ	Hualan Biological Engineering	Solid tumours	Details
ATOR-1015	ATOR-1015; ADC-1015	Phase Ⅰ	Alligator Bioscience	Solid tumours	Details
Anti-CTLA-4 antibody (Agenus)	AGEN-1181	Phase Ⅰ	Agenus	Solid tumours	Details
IDO-derived peptide vaccine/PD-L1-derived peptide vaccine	IO-102/IO-103	Phase Ⅱ	Herlev Hospital, IO Biotech	Metastatic melanoma	Details
MGD-019 (MacroGenics)	MGD-019	Phase Ⅰ	MacroGenics	Solid tumours	Details
BA-3071	BA-3071; CAB-CTLA-4	Preclinical	BioAtla, BeiGene	Cancer	Details
IO-102	IO-102	Phase Ⅱ	IO Biotech, Herlev Hospital, Merck Sharp & Dohme	Non small cell lung cancer (NSCLC), Melanoma	Details
Anti-CTLA-4 and PD-1 CAR-T cell therapy (Shanghai Cell Therapy Research Institute)		Phase Ⅱ	Shanghai Cell Therapy Research Institute	Solid tumours	Details
Ipilimumab biosimilar (Innovent Biologics)	IBI-310	Phase Ⅰ	Innovent Biologics	Solid tumours	Details
B7-2/GM-CSF cancer gene therapy	CIT	Phase Ⅰ	Radient	Solid tumours	Details
BCD-145 (Biocad)	BCD-145	Phase Ⅰ	Biocad	Metastatic melanoma	Details
Tremelimumab	CP-675; CP-675206	Phase Ⅲ	Pfizer, Amgen, MedImmune	Urogenital cancer, Small cell lung cancer (SCLC), Non small cell lung cancer (NSCLC), Mesothelioma, Solid tumours, Liver cancer, Head and neck cancer	Details
MEDI-5752	MEDI-5752	Phase Ⅰ	MedImmune	Solid tumours	Details
Zalifrelimab	AGEN-1884; RebmAb-600	Phase Ⅱ	Ludwig Institute for Cancer Research, 4-Antibody, Agenus, Recepta biopharma	Non small cell lung cancer (NSCLC), Cervical carcinoma	Details