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Purified Rat anti-Mouse GRAIL
Purified Rat anti-Mouse GRAIL
Western blot analysis of mouse GRAIL.  Lysates from four cell lines were probed with 1 µg/mL of the purified rat anti-mouse GRAIL monoclonal antibody (clone H11-744). Lane 1: NIH/3T3 retrovirally transduced with a tagged Rnf128 gene. Lane 2: NIH/3T3 retrovirally transduced with the vector alone. Lane 3: Hepa 1-6 (mouse hepatoma, ATCC CRL-1830). Lane 4: NIH/3T3 (mouse embryo fibroblasts, ATCC CRL-1658). GRAIL is identified here on transfected cells as a doublet of 75-80 kDa as opposed to the 62-66 kDa size reported to be in cells expressing GRAIL endogenously.
Purified Rat anti-Mouse GRAIL

Immunocytochemical staining of mouse GRAIL. Chamber slide cultures of NIH/3T3 retrovirally transduced with the Rnf128 gene (top row) and NIH/3T3 retrovirally transduced with vector alone (bottom row) were fixed with 2% paraformaldehyde. They were stained with either purified rat IgG2a, κ isotype control mAb (Cat. No. 559073, left column) or purified rat anti-mouse GRAIL (right column). A three-step staining procedure that employs biotin goat anti-rat IgG secondary antibody (Cat. No. 559286), streptavidin-HRP (Cat. No. 550946) and DAB (Cat. No. 550880) and hematoxylin counterstaining were used. GRAIL staining is localized in the cytoplasm of the GRAIL-expressing cells (top right panel). Original magnification: 20X.

Western blot analysis of mouse GRAIL.  Lysates from four cell lines were probed with 1 µg/mL of the purified rat anti-mouse GRAIL monoclonal antibody (clone H11-744). Lane 1: NIH/3T3 retrovirally transduced with a tagged Rnf128 gene. Lane 2: NIH/3T3 retrovirally transduced with the vector alone. Lane 3: Hepa 1-6 (mouse hepatoma, ATCC CRL-1830). Lane 4: NIH/3T3 (mouse embryo fibroblasts, ATCC CRL-1658). GRAIL is identified here on transfected cells as a doublet of 75-80 kDa as opposed to the 62-66 kDa size reported to be in cells expressing GRAIL endogenously.

Immunocytochemical staining of mouse GRAIL. Chamber slide cultures of NIH/3T3 retrovirally transduced with the Rnf128 gene (top row) and NIH/3T3 retrovirally transduced with vector alone (bottom row) were fixed with 2% paraformaldehyde. They were stained with either purified rat IgG2a, κ isotype control mAb (Cat. No. 559073, left column) or purified rat anti-mouse GRAIL (right column). A three-step staining procedure that employs biotin goat anti-rat IgG secondary antibody (Cat. No. 559286), streptavidin-HRP (Cat. No. 550946) and DAB (Cat. No. 550880) and hematoxylin counterstaining were used. GRAIL staining is localized in the cytoplasm of the GRAIL-expressing cells (top right panel). Original magnification: 20X.

製品詳細
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BD Pharmingen™
Goliath-related E3 ubiquitin-protein ligase 1, RNF128
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
Mouse GRAIL (full-length) Recombinant Protein
Western blot (Routinely Tested), (Tested During Development)
62-66 kDa
0.5 mg/ml
AB_2869097
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
557799 Rev. 7
抗体の詳細
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H11-744

The H11-744 antibody reacts with GRAIL (Gene Related to Anergy In Lymphocytes protein), a 428-amino acid glycoprotein with an apparent molecular weight of 62-66 kDa that is preferentially expressed in anergized CD4-positive CD25-negative T lymphocytes.  The GRAIL protein sequence is homologous to several zinc RING finger-containing proteins, and thus, its gene has been designated Rnf128 (RING finger protein 128).  Rnf128 transcripts are found in several mouse organs (including ovary, kidney, liver, brain, and heart), and transcription is induced rapidly (within 4 hours) in CD4-positive T cells upon in vitro induction of anergy.  The expression of GRAIL correlates with the inhibition of IL-2 production in in vitro-anergized CD4-positive CD25-negative T cells, and GRAIL expression reduces IL-2 and IL-4 production in T-cell hybridomas activated by plate-bound anti-CD3 plus anti-CD28.  GRAIL protein is localized in the recycling endosomal compartment, functions as an E3 ubiquitin ligase, and is required for anergy induction of CD4-positive T cells in vivo.

This product is sold under a license to patent number US 6,709,840.

557799 Rev. 7
フォーマットの詳細
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
557799 Rev.7
引用&参考文献
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Development References (6)

  1. Anandasabapathy N, Ford GS, Bloom D, et al. GRAIL: An E3 ubiquitin ligase that inhibits cytokine gene transcription is expressed in anergic CD4+ T cells. Immunity. 2003; 18:535-547. (Biology: Western blot). View Reference
  2. Ermann J, Szanya V, Ford GS, Paragas V, Fathman CG, Lejon K. CD4-positive CD25-positive T cells facilitate the induction of T cell anergy. J Immunol. 2001; 167:4271-4275. (Biology). View Reference
  3. Joazeiro CAP, Weissman AM. RING finger proteins: mediators of ubiquitin ligase activity. Cell. 2000; 102:549-552. (Biology). View Reference
  4. Mueller DL. E3 ubiquitin ligases as T cell anergy factors. Nat Immunol. 2004; 5(9):883-890. (Biology).
  5. Seroogy CM, Soares L, Ranheim EA, et al. The gene related to anergy in lymphocytes, an E3 ubiquitin ligase, is necessary for anergy induction in CD4 T cells. J Immunol. 2004; 173:79-85. (Biology). View Reference
  6. Soares L, Seroogy C, Skrenta H, et al. Two isoforms of otubain 1 regulate T cell anergy via GRAIL. Nat Immunol. 2004; 5:45-54. (Immunogen: Western blot). View Reference
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557799 Rev. 7

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