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HLA Class I Histocompatibility Antigen, alpha Chain G (HLAG) antibody

Details for Product No. ABIN501193
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Antigen
Synonyms MHC-G, B-F, B-F-S04, B-F-S05, B-F-S06, B-F-S07, B-FI, B-FIV, BF2, BFa2, BFw-03, BFw-05, BFz-01
Reactivity
Human
(249), (4)
Host
Mouse
(176), (76), (1)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(25), (25), (20), (9), (4), (3), (3), (3), (3), (3), (3), (3), (3), (3), (3), (3), (3), (2), (1), (1), (1)
Application
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
(161), (95), (77), (50), (47), (42), (40), (26), (24), (15), (4), (1), (1), (1)
Pubmed 4 references available
Quantity 0.1 mg
Shipping to United States (Change)
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Catalog No. ABIN501193
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Immunogen Recombinant Human HLA-G
Clone MEM-G-1
Isotype IgG1
Specificity This antibody reacts with denatured HLA-G heavy chain on Western blotting. It is reported that this monoclonal antibody can recognize all HLA-G isoforms (HLA-G1~G7). Species Reactivity: Tested: Human.
Purification Protein-A Sepharose Chromatography of hybridoma supernatant.
Alternative Name HLA class I alpha G / HLA-G
Background Human histocompatibility leukocyte antigen G (HLA-G) is an MHC class I antigen- presenting molecule. HLA-G modulates innate immunity by regulating HLA-E expression and its subsequent interaction with CD94/NKG2 receptors. HLA-G is also an effective ligand of natural killer (NK) inhibitory receptors, and it down-regulates both CD8+ and CD4+ T-cell responsiveness. HLA-G is primarily expressed on fetal cells of the human placenta, suggesting a key role for HLA-G during pregnancy in maintaining immuno- tolerance at the maternal-fetal interface.
Alternate names: HLA class I histocompatibility antigen, HLA-6.0, HLAG, MHC class I antigen G
Gene ID 3135
NCBI Accession NP_002118
UniProt P17693
Application Notes Western blotting: 1-5 µg/mL for chemiluminescence detection system. Positive Control: Human PlacentaDetailed procedure is provided in Protocols. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol SDS-PAGE & Western Blotting1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10%glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. Measure the protein concentration of the supernatant and add the cold Lysisbuffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli's sample buffer. 4) Boil the samples for 3 minutes and centrifuge. Load 10 μL of the sample per lane in a 1mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hourin a semi-dry transfer system (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for precise transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH7. 2) for 1 hour at room temperature, or overnight at 4°C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7. 2 containing 1%skimmed milk as suggest in the APPLICATIONS for 1 hour at room temperature. (Theconcentration of antibody will depend on condition. )8) Wash the membrane with PBS-T [0. 05% Tween-20 in PBS] (5 minutes x 3 times). 9) Incubate the membrane with the 1: 10,000 HRP-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at RT. 10) Wash the membrane with PBS-T (10 minutes x 3 times). 11) Wipe excess buffer on the membrane, then incubate it with appropriatechemiluminescence reagent for 1 minute. 12) Remove extra reagent from the membrane by dabbing with paper towel, and seal it inplastic wrap. 13) Expose to an X-ray film in a dark room for 3 minutes. Develop the film as usual. Thecondition for exposure and development may vary. Positive Controls: Human Placenta
Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Buffer PBS, pH 7.2 containing 50% Glycerol without preservatives.
Preservative Without preservative
Handling Advice Avoid repeated freezing and thawing.
Storage -20 °C/-80 °C
Storage Comment Store lyophilized (preferably in a desiccator) at -20°C and in aliquots at -80°C. Reconstituted antibody can be stored at 4°C for a limited period of time, it does not show decline in activity after two weeks at 4°C.
Expiry Date 12 months
Background publications Papermaster: "Introducing Paul Hargrave, the 2000 recipient of the Friedenwald award." in: Investigative ophthalmology & visual science, Vol. 42, Issue 1, pp. 1-2, 2001 (PubMed).

Menier, Saez, Horejsi et al.: "Characterization of monoclonal antibodies recognizing HLA-G or HLA-E: new tools to analyze the expression of nonclassical HLA class I molecules." in: Human immunology, Vol. 64, Issue 3, pp. 315-26, 2003 (PubMed).

Vilches: "MHC class I peptide binding and tapasin." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 171, Issue 1, pp. 3, 2003 (PubMed).

Gonen-Gross, Achdout, Arnon et al.: "The CD85J/leukocyte inhibitory receptor-1 distinguishes between conformed and beta 2-microglobulin-free HLA-G molecules." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 175, Issue 8, pp. 4866-74, 2005 (PubMed).

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