Anti-HLA-G Alexa Fluor® 488

Anti-HLA-G Alexa Fluor<sup>®</sup> 488
Regulatory status
RUO
Antigen
HLA-G
Clone
4H84
Format
Alexa Fluor® 488
Reactivity
Human
Excitation laser
blue (488 nm)
Variant
0.1 mg
A4-499-C100
In stock
374.00 USD
Variant
0.1 mg
A4-499-C100
In stock
374.00 USD
Product details
Description
Images
References
SDS download
Isotype
Mouse IgG1
Specificity
The mouse monoclonal antibody 4H84 recognizes an extracellular epitope of HLA-G molecule (39 kDa). HLA-G belongs to the MHC Class I molecules (MHC Class Ib; nonclassical) and it is expressed on the surface of trophoblast cells.
Application details
Flow cytometry: Recommended dilution: 2-5 μg/ml. Intracellular staining or surface staining after acid treatment (Polakova K. et al. Molecular immunology 1993; 1223-30).
Reactivity
Human
Immunogen
amino acids 61-83 of HLA-G of human origin
Concentration
1 mg/ml
Preparation
Purified antibody is conjugated with Alexa Fluor® 488 NHS ester under optimum conditions and unconjugated antibody and free fluorochrome are removed by size-exclusion chromatography.
Formulation
Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
Storage and handling
Store at 2-8°C. Protect from prolonged exposure to light. Do not freeze.
Exbio licence note
The product is intended For Research Use Only. Diagnostic or therapeutic applications are strictly forbidden. Products shall not be used for resale or transfer to third parties either as a stand-alone product or as a manufacture component of another product without written consent of EXBIO Praha, a.s. EXBIO Praha, a.s. will not be held responsible for patent infringement or any other violations of intellectual property rights that may occur with the use of the products. Orders for all products are accepted subject to the Term and Conditions available at www.exbio.cz. EXBIO, EXBIO Logo, and all other trademarks are property of EXBIO Praha, a.s.
Licence note
Alexa Fluor®, Pacific Blue™ and Pacific Orange™ are registered trademarks of Life Technologies Corporation.
Licence label
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.
Licence notice
This product is for research and in vitro experimental use only. It is not to be used for any other commercial purpose. Use of this product to produce products for sale or for therapeutic or drug discovery purposes is prohibited. In order to obtain a license to use this product for commercial purposes, contact The Regents of the University of California.
Antigen description
Human leukocyte antigen G (HLA-G), belonging to MHC class I glycoproteins, plays important roles in both physiological and pathological immunotolerance. It gives an inhibitory signal to cytotoxic T cells, NK cells, monocytes, and some other immune cells. It also induces regulatory T cells and anti-inflammatory macrophages. HLA-G is important e.g. for maternal tolerance to the fetus, and for immunomodulation in particular adult tissues, such as in cornea, pancreatic islets, thymus and other. On the other hand, it is expressed in many solid and hematologic malignancies, where it contributes to evasion of the immune surveillance. HLA-G expression pattern in cancer is an important prognostic factor regarding a poor clinical outcome. Unlike most other MHC glycoproteins, HLA-G acts as an immune checkpoint molecule rather than as an antigen presenting molecule. It concerns both transmembrane and soluble HLA-G isoforms. Among other, HLA-G can promote Th2 immunological response and downregulate Th1 immunological response. For its benefits regarding allograft tolerance, including embryo implantation, soluble HLA-G (sHLA-G) can be used as a marker of developmental potential of embryos during the process of in vitro fertilization. Similarly, sHLA-G concentrations in maternal serum are decreased in preeclampsia. Transplanted patients with increased sHLA-G serum levels have improved allograft acceptance. On the other hand, increased sHLA-G can also indicate presence of malignant (sometimes also of benign) tumor cells. Another important topic is induction of HLA-G expression (sometimes associated with shedding of HLA-G from the cell surface) by some anti-cancer or anti-viral therapies, which can weaken the therapy effect. Monitoring of HLA-G in patients thus has a wide usage.
Entrez Gene ID 3135
UniProt ID P17693
A4-499_FC_histogram_intra
Anti-Hu HLA-G Alexa Fluor® 488 antibody (clone 4H84) works in flow cytometry application

Analysis of the antibody staining was performed on fixed and permeabilized (FIX & PERM® Kit, ThermoFisher) cell suspension.
Anti-Hu HLA-G Alexa Fluor® 488 (concentration in sample 5 μg/ml, red-filled histogram) binds specifically to intracellular HLA-G isoforms in HLA-G5 (middle panel) and HLA-G6 (lower panel) transfected HEK-293 cells, but not to the parent HEK-293 cells (upper panel).
Level of non-specific binding was assessed using Mouse IgG1 isotype control Alexa Fluor® 488 (MOPC-21) under same conditions (concentration in sample 5 μg/ml, black-dashed histogram).
A4-499_FC_histogram_povrch
Anti-Hu HLA-G Alexa Fluor® 488 antibody (clone 4H84) works in flow cytometry application

Analysis of the antibody staining was performed on HLA-G1 transfected LCL 721.221 cells treated or untreated by short incubation with citrate-phosphate (pH 3) buffer (Polakova K. et al. Molecular immunology 1993; 1223-30). Mild acid treatment resulted in dissociation of β2-microglobulin from native HLA-G conformation.

(A) Anti-Hu HLA-G (clone 4H84) Alexa Fluor® 488 antibody (concentration in sample 5 μg/ml, red-filled histogram) binds specifically to HLA-G free heavy chain (upper panel), but not to the native HLA-G complex (lower panel).
Level of non-specific binding was assessed using Mouse IgG1 isotype control Alexa Fluor® 488 (MOPC-21) under same conditions (concentration in sample 5 μg/ml, black-dashed histogram).

(B) In the control experiment it was confirmed that Anti-Hu HLA-G (clone MEM-G/9) FITC antibody (concentration in sample 5 μg/ml, red-filled histogram) does not bind to HLA-G free heavy chain after mild acid treatment (upper panel), whereas it recognizes well native β2-microglobulin HLA-G conformation (lower panel).
Level of non-specific binding was assessed using Mouse IgG1 isotype control FITC (MOPC-21) under same conditions (concentration in sample 5 μg/ml, black-dashed histogram).

General references:

Creput C, Durrbach A, Menier C, Guettier C, Samuel D, Dausset J, Charpentier B, Carosella ED, Rouas-Freiss N. Human leukocyte antigen-G (HLA-G) expression in biliary epithelial cells is associated with allograft acceptance in liver-kidney transplantation. J Hepatol. 2003 39(4):587-94.
PubMed
Menier C, Saez B, Horejsi V, Martinozzi S, Krawice-Radanne I, Bruel S, LeDanff C, Reboul M, Hilgert I, Rabreau M, Larrad ML, Pla M, Carosella ED, Rouas-Freiss N: Characterization of monoclonal antibodies recognizing HLA-G or HLA-E: new tools to analyze the expression of nonclassical HLA class I molecules. Hum Immunol. 2003 64(3):315-26.
PubMed
Lin A, Yan WH: Heterogeneity of HLA-G expression in cancers: Facing the challenges. Front Immunol. 2018 Sep 27;9:2164.
PubMed
Hunt JS, Langat DK, McIntire RH, Morales PJ: The role of HLA-G in human pregnancy. Reprod Biol Endocrinol. 2006;4 Suppl 1(Suppl 1):S10.
PubMed
Xu HH, Yan WH, Lin A: The role of HLA-G in human papillomavirus infections and cervical carcinogenesis. Front Immunol. 2020 Jun 25;11:1349.
PubMed
Castelli EC, de Almeida BS, Muniz YC, Silva NS, Passos MR, Souza AS, Page AE, Dyble M, Smith D, Aguileta G, Bertranpetit J, Migliano AB, Duarte YA, Scliar MO, Wang J, Passos-Bueno MR, Naslavsky MS, Zatz M, Mendes CT, Donadi EA: HLA-G genetic diversity and evolutive aspects in worldwide populations. Sci Rep. 2021 Nov 29;11(1):23070.
PubMed
Curigliano G, Criscitiello C, Gelao L, Goldhirsch A: Molecular pathways: Human leukocyte antigen G (HLA-G). Clin Cancer Res. 2013 Oct 15;19(20):5564-71.
PubMed

Product specific references:

Blaschitz A, Hutter H, Dohr G: HLA Class I protein expression in the human placenta. Early Pregnancy. 2001 Jan;5(1):67-9.
PubMed
Urosevic M, Kurrer MO, Kamarashev J, Mueller B, Weder W, Burg G, Stahel RA, Dummer R, Trojan A: Human leukocyte antigen G up-regulation in lung cancer associates with high-grade histology, human leukocyte antigen class I loss and interleukin-10 production. Am J Pathol. 2001 Sep;159(3):817-24.
PubMed
Menier C, Saez B, Horejsi V, Martinozzi S, Krawice-Radanne I, Bruel S, Le Danff C, Reboul M, Hilgert I, Rabreau M, Larrad ML, Pla M, Carosella ED, Rouas-Freiss N: Characterization of monoclonal antibodies recognizing HLA-G or HLA-E: new tools to analyze the expression of nonclassical HLA class I molecules. Hum Immunol. 2003 Mar;64(3):315-26.
PubMed
Polakova K, Krcova M, Kuba D, Russ G: Analysis of HLA-G expression in malignant hematopoetic cells from leukemia patients. Leuk Res. 2003 Jul;27(7):643-8.
PubMed
Polakova K, Kuba D, Russ G: The 4H84 monoclonal antibody detecting beta2m free nonclassical HLA-G molecules also binds to free heavy chains of classical HLA class I antigens present on activated lymphocytes. Hum Immunol. 2004 Feb;65(2):157-62.
PubMed
Juch H, Blaschitz A, Daxbock C, Rueckert C, Kofler K, Dohr G: A novel sandwich ELISA for alpha1 domain based detection of soluble HLA-G heavy chains. J Immunol Methods. 2005 Dec 20;307(1-2):96-106.
PubMed
Barrier BF, Kendall BS, Sharpe-Timms KL, Kost ER: Characterization of human leukocyte antigen-G (HLA-G) expression in endometrial adenocarcinoma. Gynecol Oncol. 2006 Oct;103(1):25-30.
PubMed
Paul P, Rouas-Freiss N, Khalil-Daher I, Moreau P, Riteau B, Le Gal FA, Avril MF, Dausset J, Guillet JG, Carosella ED: HLA-G expression in melanoma: a way for tumor cells to escape from immunosurveillance. Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4510-5.
PubMed
Gonen-Gross T, Achdout H, Arnon TI, Gazit R, Stern N, Horejsí V, Goldman-Wohl D, Yagel S, Mandelboim O: The CD85J/leukocyte inhibitory receptor-1 distinguishes between conformed and beta 2-microglobulin-free HLA-G molecules. J Immunol. 2005 Oct 15;175(8):4866-74.
PubMed
Creput C, Le Friec G, Bahri R, Amiot L, Charpentier B, Carosella E, Rouas-Freiss N, Durrbach A: Detection of HLA-G in serum and graft biopsy associated with fewer acute rejections following combined liver-kidney transplantation: Possible implications for monitoring patients. Hum Immunol. 2003 Nov;64(11):1033-8.
PubMed
Poláková K, Bennink JR, W Yewdell J, Bystrická M, Bandzuchová E, Russ G: Mild acid treatment induces cross-reactivity of 4H84 monoclonal antibody specific to nonclassical HLA-G antigen with classical HLA class I molecules. Hum Immunol. 2003 Feb;64(2):256-64.
PubMed
Zhang X, Lin A, Han QY, Zhang JG, Chen QY, Ye YH, Zhou WJ, Xu HH, Gan J, Yan WH: Intratumor heterogeneity of HLA-G expression in cancer lesions. Front Immunol. 2020 Nov 19;11:565759.
PubMed
Zhao L, Teklemariam T, Hantash BM: Reassessment of HLA-G isoform specificity of MEM-G/9 and 4H84 monoclonal antibodies. Tissue Antigens 2012 Sep;80(3):231-8.
PubMed
Variant
0.1 mg
A4-499-C100
In stock
374.00 USD
Variant
0.1 mg
A4-499-C100
In stock
374.00 USD

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