PrimePCR™ Probe Assay: HLA-B, Human

RT

Real-time PCR probe assay designed for gene expression analysis. Probe assays consist of unlabeled PCR primers and a dual labeled fluorescent probe.

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Assay Information

Technology:   qPCR
Assay Type:   Probe
Application:   Gene Expression
Unique Assay ID:   qHsaCEP0040193
Assay Design:   exonic
Chromosome Location:   6:31321920-31322010question
Amplicon Length:   61
Splice Variants Targeted:   ENST00000383487 ENST00000383483 ENST00000458668 ENST00000400341 ENST00000437265 ENST00000421349 ENST00000553172 ENST00000376228 ENST00000383329 ENST00000539307 ENST00000453809 ENST00000414249 ENST00000419590 ENST00000438171 ENST00000552865 ENST00000412585 ENST00000436729 ENST00000435618 ENST00000546599 ENST00000419135 ENST00000433153 ENST00000429840 ENST00000457903 ENST00000425798 ENST00000450871 ENST00000458192 ENST00000422921 ENST00000456487 ENST00000420206 ENST00000423509 ENST00000430940 ENST

Gene Information

HLA-B belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta-2 microglobulin). The heavy chain is anchored in the membrane. Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. They are expressed in nearly all cells. The heavy chain is approximately 45 kDa and its gene contains 8 exons. Exon 1 encodes the leader peptide exon 2 and 3 encode the alpha1 and alpha2 domains which both bind the peptide exon 4 encodes the alpha3 domain exon 5 encodes the transmembrane region and exons 6 and 7 encode the cytoplasmic tail. Polymorphisms within exon 2 and exon 3 are responsible for the peptide binding specificity of each class one molecule. Typing for these polymorphisms is routinely done for bone marrow and kidney transplantation. Hundreds of HLA-B alleles have been described. [provided by RefSeq Jul 2008]

Gene Symbol:   HLA-B
Gene Name:   major histocompatibility complex, class I, B
Aliases:   AS, Bw-45, Bw-48, Bw-56, HLA-B-5601, HLA-B-6701, HLA-B27, HLA-B73, HLAB, MGC111087, SPDA1
RefSeq:   NC_000006.11 NG_002397.2 NG_023187.1 NT_007592.15 NT_113891.2 NT_167246.1 NT_167247.1 NT_167248.1 NT_167249.1
Ensembl:   ENSG00000234745
Entrez:   3106
UniGene:   Hs.77961
Chromosome Mapping:   6p21.3

The below validation information is for the Primer Pair only   Download Validation Data (.pdf)


Products used to generate validation data:

Real-Time PCR Instrument CFX384 Real-Time PCR Detection System
Reverse Transcription Reagent iScript™ Advanced cDNA Synthesis Kit for RT-qPCR
Real-Time PCR Supermix SsoAdvanced™ SYBR® Green Supermix
Experimental Sample qPCR Human Reference Total RNA

Summary Data:

R2 0.999800
y-intercept 39.380000
Efficiency 87

Amplification Plot
Amplification of cDNA generated from universal RNA.

Amplification of cDNA generated from universal RNA.

Melt Peak
Melt curve analysis of above amplification.

Melt curve analysis of above amplification.

Standard Curve
Standard curve generated using 20 million copies of template diluted 10 fold to 20 copies.

Standard curve generated using 20 million copies of template diluted 10 fold to 20 copies.

Number Description Download
6262 PrimePCR Assays: Meeting the MIQE Guidelines by Full Wet-Lab Validation, Rev B Click to download
6263 PrimePCR Pathway Analysis: Pathway Curation and Real-Time PCR Panel Design Strategy, Rev B Click to download
6290 PrimePCR Assays and Panels Brochure, Rev D Click to download
LIT10026370 Instruction Manual, PrimePCR Assays, Panels, and Controls, Rev E Click to download
10039761 PrimePCR™ Assays Quick Guide, Ver B Click to download