製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to Gephyrin
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: ICC/IF, IHC-FoFr, WBmore details
  • 種交差性
    交差種: Mouse, Rat
    交差が予測される動物種: Chicken, Human, Xenopus laevis
  • 免疫原

    Synthetic peptide within Mouse Gephyrin aa 700 to the C-terminus conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    Database link: Q8BUV3
    (Peptide available as ab32205)

  • ポジティブ・コントロール
    • WB: Mouse brain whole cell lysate. NIH/3T3, MEF1 and PC-12 whole cell lysate. Mouse liver, heart, skeletal muscle and kidney tissue lysate; Human pancreas tissue lysate; Rat liver, kidney and heart tissue lysate. IHC-PFA perfusion fixed frozen: Rat brain cortex tissue. ICC/IF: MEF1 and PC-12 cells.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab32206 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 1 µg/ml.
IHC-FoFr 1/1000.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 93,75,54 kDa (predicted molecular weight: 83 kDa).

The 93 kDa band corresponds to Isoform 1 of Gephyrin as observed in the literature. We suspect that the 75 kDa band corresponds to another isoform of Gephyrin. The identity of the 54 kDa band is unknown. All three of the bands we observe on WB are partially blocked by addition of the immunising peptide (ab32205).

ターゲット情報

  • 機能
    Microtubule-associated protein involved in membrane protein-cytoskeleton interactions. It is thought to anchor the inhibitory glycine receptor (GLYR) to subsynaptic microtubules (By similarity). Catalyzes two steps in the biosynthesis of the molybdenum cofactor. In the first step, molybdopterin is adenylated. Subsequently, molybdate is inserted into adenylated molybdopterin and AMP is released.
  • パスウェイ
    Cofactor biosynthesis; molybdopterin biosynthesis.
  • 関連疾患
    Defects in GPHN are the cause of molybdenum cofactor deficiency type C (MOCOD type C) [MIM:252150]. MOCOD type C is an autosomal recessive disease which leads to the pleiotropic loss of all molybdoenzyme activities and is characterized by severe neurological damage, neonatal seizures and early childhood death.
    Defects in GPHN are a cause of startle disease (STHE) [MIM:149400]; also known as hyperekplexia. STHE is a genetically heterogeneous neurologic disorder characterized by muscular rigidity of central nervous system origin, particularly in the neonatal period, and by an exaggerated startle response to unexpected acoustic or tactile stimuli.
  • 配列類似性
    In the N-terminal section; belongs to the moaB/mog family.
    In the C-terminal section; belongs to the moeA family.
  • 細胞内局在
    Cell junction > synapse. Cell junction > synapse > postsynaptic cell membrane. Cytoplasm > cytoskeleton. Cytoplasmic face of glycinergic postsynaptic membranes.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Domain E antibody
    • Domain G antibody
    • GEPH antibody
    • GEPH_HUMAN antibody
    • GPH antibody
    • GPHN antibody
    • GPHRYN antibody
    • KIAA1385 antibody
    • Molybdopterin molybdenumtransferase antibody
    • MPT adenylyltransferase antibody
    • MPT Mo-transferase antibody
    see all

画像

  • Immunofluorescent staining for Gephyrin in rat brain cortex using Rabbit polyclonal to Gephyrin (ab32206; 1/1000). The staining is located in the cytoplasm and in some of the processes of cortical neurons. This staining is observed in many other brain areas. The picture was acquired using a X20 objective. Protocol details: Rats were intracardially perfused with 4% paraformaldehyde. Whole brain tissue was post-fixed overnight in the same fixative, and cryoprotected in 20% sucrose and frozen in OCT. 30 µm coronal sections were cut by cyrostat for use in fre floating IHC. Primary antibody ab32206 was incubated overnight at 1/1000 at room temperature. Secondary antibody Alexa fluor® 488 1/1000 was incubated for 2 hours at room temperature.

  • All lanes : Anti-Gephyrin antibody (ab32206) at 1 µg/ml

    Lane 1 : Adult mouse brain whole cell lysate
    Lane 2 : Mouse brain tissue lysate - total protein (0 days) (ab7188) at 20 µg
    Lane 3 : Adult mouse brain whole cell lysate at 20 µg with Mouse Gephyrin peptide (ab32205) at 1 µg/ml
    Lane 4 : Mouse brain tissue lysate - total protein (0 days) (ab7188) at 20 µg with Mouse Gephyrin peptide (ab32205) at 1 µg/ml

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 83 kDa



    The 93 kDa band corresponds to Isoform 1 of Gephyrin as observed in the literature. We suspect that the 75 kDa band corresponds to another isoform of Gephyrin. The identity of the 54 kDa band is unknown. All three of the bands we observe on WB are partially blocked by addition of the immunising peptide (ab32205).

  • All lanes : Anti-Gephyrin antibody (ab32206) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 3 : Mouse liver tissue lysate - normal tissue
    Lane 4 : Mouse heart tissue lysate
    Lane 5 : Mouse kidney tissue lysate
    Lane 6 : Mouse pancreas tissue lysate - total protein (ab29363)
    Lane 7 : Mouse skeletal muscle tissue lysate - total protein (ab29711)
    Lane 8 : PC-12 (Rat adrenal pheochromocytoma cell line) whole cell lysate
    Lane 9 : Rat liver tissue lysate
    Lane 10 : Rat heart tissue lysate
    Lane 11 : Kidney (Rat) Whole Cell Lysate - normal tissue (ab29480)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 83 kDa
    Observed band size: 75,93 kDa (why is the actual band size different from the predicted?)

  • ICC/IF image of ab32206 stained MEF1 (Mouse embryonic fibroblast cell line) cells. The cells were 4% PFA fixed (10 minutes), permeabilized in 0.1% PBS-Tween (20 minutes) and incubated with the antibody (ab32206, 1 µg/ml) for 1 hour at room temperature. 1% BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • ICC/IF image of ab32206 stained PC-12 (Rat adrenal gland pheochromocytoma cell line) cells. The cells were fixed in 100% methanol for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32206, 1 µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

参考文献

This product has been referenced in:
  • Wang S  et al. Macroglia-derived thrombospondin 2 regulates alterations of presynaptic proteins of retinal neurons following elevated hydrostatic pressure. PLoS One 12:e0185388 (2017). Read more (PubMed: 28953973) »
  • Nunez-Parra A  et al. Disruption of centrifugal inhibition to olfactory bulb granule cells impairs olfactory discrimination. Proc Natl Acad Sci U S A 110:14777-82 (2013). Read more (PubMed: 23959889) »

See all 9 Publications for this product

レビューと Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (cerebral organoids derived from human induced plur)
Permeabilization
No
Specification
cerebral organoids derived from human induced plur
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Mar 13 2017

Application
Western blot
Loading amount
2 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Xenopus laevis Cell lysate - whole cell (Tectal cell)
Specification
Tectal cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Jul 31 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Rat brain)
Specification
Rat brain
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Username

Dr. Sophie Pezet

Verified customer

投稿 Jan 16 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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