HLA-DQA2 Protein (AA 24-255) (rho-1D4 tag)

Catalog No. ABIN3092193

Product Details

Target: HLA-DQA2 (Major Histocompatibility Complex, Class II, DQ alpha 2 (HLA-DQA2))

Protein Type: Recombinant

Protein Characteristics: AA 24-255

Origin: Human

Source: Insect Cells

Purification tag / Conjugate: This HLA-DQA2 protein is labelled with rho-1D4 tag.

Application: SDS-PAGE (SDS), Crystallization (Crys), ELISA, Western Blotting (WB)

Sequence: EDIVADHVAS YGVNFYQSHG PSGQYTHEFD GDEEFYVDLE TKETVWQLPM FSKFISFDPQ SALRNMAVGK HTLEFMMRQS NSTAATNEVP EVTVFSKFPV TLGQPNTLIC LVDNIFPPVV NITWLSNGHS VTEGVSETSF LSKSDHSFFK ISYLTFLPSA DEIYDCKVEH WGLDEPLLKH WEPEIPAPMS ELTETLVCAL GLSVGLMGIV VGTVFIIQGL RSVGASRHQG LL
Sequence without tag. Tag location is at the discretion of the manufacturer. If you have a special request, please contact us.

Characteristics:

• Made in Germany - from design to production - by highly experienced protein experts.
• Human HLA-DQA2 Protein (raised in Insect Cells) purified by multi-step, protein-specific process to ensure crystallization grade.
• State-of-the-art algorithm used for plasmid design (Gene synthesis).

This protein is a made to order protein and will be made for the first time for your order. Our experts in the lab will ensure that you receive a correctly folded protein.

The big advantage of ordering our made-to-order proteins in comparison to ordering custom made proteins from other companies is that there is no financial obligation in case the protein cannot be expressed or purified.

In the unlikely event that the protein cannot be expressed or purified we do not charge anything (other companies might charge you for any performed steps in the expression process for custom-made proteins, e.g. fees might apply for the expression plasmid, the first expression experiments or purification optimization).

When you order this made-to-order protein you will only pay upon receival of the correctly folded protein. With no financial risk on your end you can rest assured that our experienced protein experts will do everything to make sure that you receive the protein you ordered.

The concentration of our recombinant proteins is measured using the absorbance at 280nm. The protein's absorbance will be measured in several dilutions and is measured against its specific reference buffer.

The concentration of the protein is calculated using its specific absorption coefficient. We use the Expasy's protparam tool to determine the absorption coefficient of each protein.

Purification: Three step purification of membrane proteins expressed in baculovirus infected SF9 insect cells:

1. Membrane proteins are fractioned by ultracentrifugation and subsequently solubilized with different detergents (detergent screen). Samples are analyzed by Western blot.
2. The best performing detergent is used for solubilization and the proteins are purified via their rho1D4 tag via two rho1D4 antibody columns: one DTT resistant, the other one not. Eluate fractions are analyzed by Western blot.
3. Protein containing fractions of the best purification are subjected to second purification step through size exclusion chromatograph. Eluate fractions are analyzed by SDS-PAGE and Western blot.

Purity: >95 % as determined by SDS PAGE, Size Exclusion Chromatography and Western Blot.

Sterility: 0.22 μm filtered

Endotoxin Level: Protein is endotoxin-free.

Grade: Crystallography grade

Application Details

Application Notes: In addition to the applications listed above we expect the protein to work for functional studies as well. As the protein has not been tested for functional studies yet we cannot offer a gurantee though.

Comment:

In cases in which it is highly likely that the recombinant protein with the default tag will be insoluble our protein lab may suggest a higher molecular weight tag (e.g. GST-tag) instead to increase solubility. We will discuss all possible options with you in detail to assure that you receive your protein of interest.

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: 100 mM NaCL, 20 mM Hepes, 10% glycerol. pH value is at the discretion of the manufacturer.

Handling Advice: Avoid repeated freeze-thaw cycles.

Storage: -80 °C

Storage Comment: Store at -80°C.

Expiry Date: Unlimited (if stored properly)

Target Details

Target: HLA-DQA2 (Major Histocompatibility Complex, Class II, DQ alpha 2 (HLA-DQA2))

Alternative Name: HLA-DQA2 (HLA-DQA2 Products)

Synonyms: DX-ALPHA Protein, HLA-DXA Protein, major histocompatibility complex, class II, DQ alpha 2 Protein, HLA-DQA2 Protein

Background: Binds peptides derived from antigens that access the endocytic route of antigen presenting cells (APC) and presents them on the cell surface for recognition by the CD4 T-cells. The peptide binding cleft accommodates peptides of 10-30 residues. The peptides presented by MHC class II molecules are generated mostly by degradation of proteins that access the endocytic route, where they are processed by lysosomal proteases and other hydrolases. Exogenous antigens that have been endocytosed by the APC are thus readily available for presentation via MHC II molecules, and for this reason this antigen presentation pathway is usually referred to as exogenous. As membrane proteins on their way to degradation in lysosomes as part of their normal turn-over are also contained in the endosomal/lysosomal compartments, exogenous antigens must compete with those derived from endogenous components. Autophagy is also a source of endogenous peptides, autophagosomes constitutively fuse with MHC class II loading compartments. In addition to APCs, other cells of the gastrointestinal tract, such as epithelial cells, express MHC class II molecules and CD74 and act as APCs, which is an unusual trait of the GI tract. To produce a MHC class II molecule that presents an antigen, three MHC class II molecules (heterodimers of an alpha and a beta chain) associate with a CD74 trimer in the ER to form a heterononamer. Soon after the entry of this complex into the endosomal/lysosomal system where antigen processing occurs, CD74 undergoes a sequential degradation by various proteases, including CTSS and CTSL, leaving a small fragment termed CLIP (class-II-associated invariant chain peptide). The removal of CLIP is facilitated by HLA-DM via direct binding to the alpha-beta-CLIP complex so that CLIP is released. HLA-DM stabilizes MHC class II molecules until primary high affinity antigenic peptides are bound. The MHC II molecule bound to a peptide is then transported to the cell membrane surface. In B-cells, the interaction between HLA-DM and MHC class II molecules is regulated by HLA-DO. Primary dendritic cells (DCs) also to express HLA-DO. Lysosomal microenvironment has been implicated in the regulation of antigen loading into MHC II molecules, increased acidification produces increased proteolysis and efficient peptide loading. {ECO:0000269|PubMed:22407913}.

Molecular Weight: 27.0 kDa Including tag.

UniProt: P01906

Pathways: TCR Signaling