Recombinant Human CD38 His-tag Avi-tag Protein, CF

Catalog #: AVI2404 Datasheet / COA / SDS

Biotinylated

Catalog #	Availability	Size / Price	Qty
AVI2404-050

Recombinant Human CD38 His-tag Avi-tag Protein Binding Activity

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human CD38 His-tag Avi-tag Protein, CF Summary

Learn more about Avi-tag Biotinylated Proteins

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its ability to convert the substrate nicotinamide guanine dinucleotide (NGD⁺) to cyclic GDP-ribose. Graeff, R.M. et al. (1994) J. Biol. Chem. 269:30260. The specific activity is >1500 pmol/min/μg, as measured under the described conditions. Measured by its binding ability in a functional ELISA. When Human CD38 Antibody (Catalog # AF2404) is immobilized at 1 µg/mL (100 µL/well), it binds to Biotinylated Recombinant Human CD38 His-tag Avi-tag (Catalog # AVI2404) with an ED₅₀ of 1-6 ng/mL.

Source

Human embryonic kidney cell, HEK293-derived human CD38 protein

Human CD38 (Val43-Ile300) Accession # P28907.2	HHHHHH	Avi-tag
N-terminus		C-terminus

N-terminal Sequence
Analysis

Val43

Structure / Form

Biotinylated via Avi-tag

Predicted Molecular Mass

33 kDa

SDS-PAGE

40-52 kDa, under reducing conditions

Product Datasheets

AVI2404

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AVI2404

Formulation	Supplied as a 0.2 μm filtered solution in PBS.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 6 months from date of receipt, -20 to -70 °C as supplied. 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials

Assay Buffer: 50 mM MES, pH 6.5
Recombinant Human CD38 His-tag Avi-tag (rhCD38/His/Avi-tag) (Catalog # AVI2404)
Substrate: Nicotinamide guanine dinucleotide (NGD⁺) (Sigma, Catalog # N5131), 10 mM stock in deionized water
F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

Dilute rhCD38/His/Avi-tag to 2 ng/µL in Assay Buffer.
Dilute Substrate to 200 µM in Assay Buffer.
Load in plate, 50 µL of 2 ng/µL rhCD38/His/Avi-tag, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
Read at excitation and emission wavelengths of 300 nm and 410 nm (top read), respectively in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (RFU/min) x Conversion Factor** (pmol/RFU)
	amount of enzyme (µg)

*Adjusted for Substrate Blank
**Derived using calibration standard cyclic GDP ribose (cGDPR).

Per Well:

rhCD38/His/Avi-tag: 0.1 µg
Substrate: 100 µM

Scientific Data

Binding Activity

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When Human CD38 Antibody (AF2404) is immobilized at 1 μg/mL (100 μL/well), it binds to Biotinylated Recombinant Human CD38 His-tag Avi-tag (AVI2404) with an ED₅₀ of 1-6 ng/mL.

SDS-PAGE

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2 μg/lane of Recombinant Human CD38 His-tag Avi-tag Protein (AVI2404) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 40-52 kDa.

Reconstitution Calculator

Background: CD38

CD38, also known as ADP-ribosyl cyclase, is a Type II integral membrane protein. The enzyme is able to transform NAD(P)+ into three different products with calcium mobilizing ability, cyclic ADP-ribose, NAADP+, and ADP-ribose (1). CD38 is expressed in B and T lymphocytes, osteoclasts, and in cardiac, pancreatic, liver and kidney cells (2, 3). Through its production of cyclic ADP-ribose, CD38 modulates calcium-mediated signal transduction in many types of cells, including neutrophils and pancreatic beta cells (4, 5). Our Avi-tag Biotinylated Recombinant Human CD38 features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

References

Schuber, F. and F.E. Lund (2004) Curr. Mol. Med. 4:249.
Jackson, D.G. and J.I. Bell (1990) J. Immunol. 144:2811.
Sun, L. et al. (1999) J. Cell Biol. 146:1161.
Partida-Sanchez, S. et al. (2001) Nature Med. 7:1209.
Kato, I. et al. (1995) J. Biol. Chem. 270:30045.

Long Name

Cluster of Differentiation 38

Entrez Gene IDs

952 (Human); 12494 (Mouse); 25668 (Rat); 102126394 (Cynomolgus Monkey)

Alternate Names

2'-phospho-ADP-ribosyl cyclase; 2'-phospho-ADP-ribosyl cyclase/2'-phospho-cyclic-ADP-ribose transferase (EC:2.4.99.20); 2'-phospho-cyclic-ADP-ribose transferase; ADPRC 1; ADPRC1; ADP-ribosyl cyclase 1; ADP-ribosyl Cyclase; ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1; ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase; cADPr hydrolase 1; CD_antigen: CD38; CD38 antigen (p45); CD38 antigen; CD38 molecule; CD38; cluster of differentiation 38; Cyclic ADP-ribose hydrolase 1; Cyclic ADP-ribose Hydrolase; EC 2.4.99.20; EC 3.2.2.6; EC:3.2.2.6; NAD(+) nucleosidase; T10