Human leukocyte antigen G,HLA-G ELISA Kit

Instructions
Code CSB-E09401h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human leukocyte antigen G (HLA-G) ELISA Kit is specifically designed to standardize the detection of HLA-G in serum, plasma, or tissue homogenates. It helps to streamline the experiment and obtain ideal results due to its excellent performances in specificity, sensitivity, precision, linearity, recovery, and lot-to-lot consistency. The assay combines the sandwich-ELISA mechanism with enzyme-substrate chromogenic reaction to complete the detection. The solution color develops in proportion to the HLA-G concentrations in the sample. And the color intensity can be measured at 450 nm via a microplate reader.

HLA-G is a non-classical HLA class I antigen with diverse protein isoforms. HLA-G acts as an immune-modulatory molecule and plays an important role in fetal-maternal immune tolerance during pregnancy by interacting with immune cells of both innate and adaptive responses. It is restricted to several healthy adult tissues, including the extravillous cytotrophoblasts, thymic medulla, cornea, and pancreatic islets. However, HLA-G expression is found in various pathological conditions such as cancers, organ transplantation, viral infection, and autoimmune and inflammatory diseases. The expression of HLA-G in cancers is highly related to immune inhibitory microenvironments, late tumor stage, and poor therapeutic responses and prognosis.

Target Name major histocompatibility complex, class I, G
Alternative Names B2 microglobulin ELISA Kit; DADB-15K14.8 ELISA Kit; HLA 6.0 ELISA Kit; HLA class I histocompatibility antigen alpha chain G ELISA Kit; HLA class I histocompatibility antigen; alpha chain G ELISA Kit; HLA class I molecule ELISA Kit; HLA G ELISA Kit; HLA G antigen ELISA Kit; HLA G histocompatibility antigen class I G ELISA Kit; HLA G3 ELISA Kit; HLA-G ELISA Kit; HLA-G histocompatibility antigen; class I ELISA Kit; HLA60 ELISA Kit; HLAG ELISA Kit; HLAG_HUMAN ELISA Kit; Major histocompatibility complex class I G ELISA Kit; MHC class I antigen ELISA Kit; MHC class I antigen G ELISA Kit; MHC G ELISA Kit; T-cell A locus ELISA Kit; TCA ELISA Kit
Abbreviation HLA-G
Uniprot No. P17693
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
Detection Range 1.25 ng/mL-80 ng/mL
Sensitivity 0.31 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Immunology
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human HLA-G in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:1 Average % 93  
Range % 86-98  
1:2 Average % 103  
Range % 97-108  
1:4 Average % 93  
Range % 85-98  
1:8 Average % 97  
Range % 91-101  
Recovery
The recovery of human HLA-G spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 86 80-94  
EDTA plasma (n=4) 96 92-104  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average Corrected  
80 2.961 2.960 2.961 2.828  
40 2.252 2.205 2.229 2.096  
20 1.266 1.286 1.276 1.143  
10 0.731 0.755 0.743 0.610  
5 0.368 0.400 0.384 0.251  
2.5 0.302 0.322 0.312 0.179  
1.25 0.227 0.215 0.221 0.088  
0 0.134 0.132 0.133    
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human HLA-G antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled HLA-G antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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Target Data

Function Involved in the presentation of foreign antigens to the immune system. Plays a role in maternal tolerance of the fetus by mediating protection from the deleterious effects of natural killer cells, cytotoxic T-lymphocytes, macrophages and mononuclear cells.
Gene References into Functions
  1. HLA-G, NRP1, and PD-1, may be involved in the immune response in psoriatic patients PMID: 29790686
  2. Considering that others 14-bp associations were inconclusive and that other variation sites observed at HLA-G 3'UTR exhibit a proven role on post-transcriptional regulation of HLA-G expression, the complete 3'UTR segment should be analyzed in terms of disease susceptibility, instead of a single polymorphism. PMID: 30102938
  3. study demonstrated that HLA-G 14-bp Ins/Del polymorphism may exert no influence on susceptibility to viruses - Meta-analysis PMID: 30235670
  4. The findings indicate that GPER/miR148a/HLAG signaling pathway may mediates the development of ovarian endometriosis and may become a potential therapeutic target for the treatment of endometriosis. PMID: 29845209
  5. Significantly different distribution of HLA-G polymorphism (rs1611715), but not the serum level of sHLA-G, were found between multiple sclerosis patients and healthy individuals. PMID: 29924453
  6. These results indicate the importance of the HLA-G promoter SNPs in the pregnancy outcome. But to reach a more definite conclusion, subsequent studies on 3' UTR and other positions with polymorphism in the 5' UTR regions larger samples are necessary. PMID: 29797531
  7. HLA-G expression is associated with poor survival in stage III gastric cancer patients and represents a possible immunoescape mechanism of cancer cells. PMID: 29845360
  8. this study shows that HLA-G molecules have a role in predicting the newborn's likelihood for oral HPV infection at birth. PMID: 29544814
  9. HLA-G is highly represented in ovarian carcinoma suggesting a potential association with progressive disease mechanism PMID: 29499226
  10. this study shows that HLA-G is expressed in intestinal samples of ulcerative colitis and Crohn's disease patients PMID: 29588183
  11. this study shows that HLA-G mediated immune regulation is impaired by a single amino acid exchange in the alpha 2 domain PMID: 29605689
  12. No significant differences in single nucleotide polymorphisms in the 3' untranslated region of HLA-G in placentas between spontaneous preterm birth and preeclampsia. PMID: 29540242
  13. the HLA-G 14bp ins/del gene polymorphism is an important risk factor for the incidence and poor outcome of NHL cases. PMID: 29388862
  14. these findings showed that HLA-G overexpression in tumor tissue correlated with poor prognosis in pancreatic adenocarcinoma PMID: 29549230
  15. This meta-analysis suggested that the HLA-G 14-bp insertion allele may increase the risk of recurrent implantation failure in Caucasians. PMID: 28707147
  16. Study shows that HLA-G alleles are skewed in autism spectrum disorder (ASD) children. HLA-G alleles seen in ASD result in immune activation. The skewing of HLA-G alleles seen in ASD might play a pathogenic role. PMID: 28923404
  17. these data confirm an important role of HLA-G 3'UTR polymorphisms in the development of severe forms of sepsis in Brazil PMID: 28941746
  18. a 14bp indel in HLA-G associated with increased flow-mediated dilatation of the brachial artery in renal transplant recipients PMID: 28987961
  19. the maternal HLA-G 14-bp Ins/Del polymorphism is not associated with preeclampsia risk. PMID: 29105301
  20. the HLA-G 14-bp Ins/Del polymorphism may be a marker for genetic susceptibility to chronic hepatitis B infection. PMID: 28929613
  21. The presence of the HLA-G +3142 CC genotype was associated with higher susceptibility to CMV infection after kidney transplantation. PMID: 29113092
  22. The results from the study suggest a possible involvement of HLA-G in the risk modulation toward hepatitis C virus infection. PMID: 28083985
  23. our results suggest that elevated levels of HLA- G expression, precisely sHLA- G1 homodimers, are indeed associated with persistent infection with HBV. sHLA- G (sHLA- G1 and HLA- G5 both) levels may be a prognostic indicator for spontaneous recov-ery. PMID: 28429836
  24. Results show that HLA-G rs1233334:CT protected against progression of endometriosis. PMID: 29234882
  25. The results suggest that HLA-G polymorphism has small effect on systemic lupus erythematosus susceptibility and that soluble HLA-G may be involved in the pathogenesis of the disease. PMID: 28695673
  26. sHLA-G levels is an independent prognosis factor and improves the prognostic stratification offered by traditional prognosticators in CRC patients. PMID: 28415627
  27. we found a significant association between HLA-G rs1063320 (thorn3142G>C) and 14-bp ins/del variants and risk of recurrent spontaneous abortion. PMID: 28600033
  28. HLA-G del/del genotypes were more frequent among mothers with gestational diabetes than in control. Babies carrying HLA-G del/del showed the highest sHLA-G levels. PMID: 28655969
  29. No statistically significant differences in either HLA-G allele or genotype frequencies between pre-eclampsia cases and control group have been observed. PMID: 29061057
  30. Overexpressed HLA-G revealed their ability to inhibit the cell proliferation and cytotoxic activity of NK-92MI cells, and reduce the secretion of IFN-gamma and TNF-alpha through immunoglobulin-like transcript 2. PMID: 29224003
  31. Soluble HLA-G was significantly higher in the persistent wheezing (positive modified asthma predictive index) infant group compared with the transient wheezing (negative modified asthma predictive index) group (P = 0.008). PMID: 27880031
  32. this study shows associations between fetal HLA-G genotype and birth weight and placental weight in a large cohort of pregnant women PMID: 28267635
  33. evidence is presented that HLA-G1 monomer exhibits the therapeutic effects on atopic dermatitis-like skin lesions in mice PMID: 28675838
  34. this study shows that HLA-G is expressed by ARPE-19 retinal pigment epithelium cells and is upregulated as a response to pro-inflammatory cytokines PMID: 28442288
  35. Cellular activation and pathological processes are associated with an aberrant or a neo-expression of HLA-G/soluble HLA-G. (Review) PMID: 27440734
  36. the +3142 C>G, but not the 14bp INS/DEL polymorphism may constitute a genetic susceptibility factor to multiple sclerosis in the Tunisian population. PMID: 27771469
  37. novel HLA-G-regulating miRs, miR-628-5p and miR-548q, were identified in renal cell carcinoma PMID: 27057628
  38. high expression of total soluble HLA-G and vesicular soluble HLA-G, together with the presence of the 14-bp deletion allele, might be involved in implantation failure PMID: 26959830
  39. the presented data suggest that the investigated HLA-G*0105N allele is potentially associated with recurrent spontaneous abortions through linkage disequilibrium with other genetic elements. PMID: 26754761
  40. these findings demonstrate the association of HLA-G polymorphism with breast cancer susceptibility in Tunisian population PMID: 26754763
  41. results strongly suggest the involvement of HLA-G in human African trypanosomiasis disease progression PMID: 27470243
  42. women with preterm premature rupture of membranes had significantly higher serum and vaginal sHLA-G concentrations compared to controls PMID: 27232355
  43. Serum levels of soluble HLA-G were similar in CMV- and CMV+ subjects but levels in culture supernatants were significantly higher in cells from CMV+ than from CMV- subjects stimulated with CMV antigens. The HLA-G ligand KIR2DL4 was mainly expressed on NK cells and CD56+ T cells with no differences between CMV+ and CMV- subjects. PMID: 28817184
  44. our study is the first to suggest that HLA-G expression may influence the clinical outcome of chronic myeloid leukemia PMID: 28841441
  45. SV40 immune-inhibitory direct effect and the presence of high levels of the immune-inhibitory HLA-G molecules could co-operate in impairing B lymphocyte activation towards SV40 specific peptides. PMID: 27443345
  46. HLA-G 3'UTR polymorphisms associated with a greater magnitude of HLA-G production were associated with differentiated thyroid tumours and with variables implicated in poor prognosis. PMID: 27914217
  47. The inhibitory properties of bronchial endothelial cellss are dysregulated in stable lung transplant recipients via TGF-beta, IL-10 and HLA-G signaling pathway. PMID: 27820781
  48. HLA-G expression was upregulated in chondrocyte-differentiated mesenchymal stem cells under hypoxia context and could be boosted in allogenic settings. PMID: 27465875
  49. this review provides new insights into the mechanisms controlling the expression and function of HLA-G at the maternal-fetal interface, and discuss their relevance for fetal tolerance PMID: 28279591
  50. we propose a massively parallel sequencing (NGS) with a bioinformatics strategy to evaluate the entire HLA-G regulatory and coding segments PMID: 28135606

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Subcellular Location Isoform 1: Cell membrane, Single-pass type I membrane protein, SUBCELLULAR LOCATION: Isoform 2: Cell membrane, Single-pass type I membrane protein, SUBCELLULAR LOCATION: Isoform 3: Cell membrane, Single-pass type I membrane protein, SUBCELLULAR LOCATION: Isoform 4: Cell membrane, Single-pass type I membrane protein, SUBCELLULAR LOCATION: Isoform 5: Secreted, SUBCELLULAR LOCATION: Isoform 6: Secreted, SUBCELLULAR LOCATION: Isoform 7: Secreted
Protein Families MHC class I family
Tissue Specificity Expressed in trophoblasts. Expressed in fetal eye and thymus (PubMed:2336406). Also expressed in adult eye (PubMed:1570318). Isoform 4: Expressed in fetal first trimester trophoblasts (PubMed:7589701). Isoform 7: Expressed in first trimester, second trime
Database Links

HGNC: 4964

OMIM: 142871

KEGG: hsa:3135

STRING: 9606.ENSP00000353472

UniGene: Hs.512152

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