Overview

  • Product nameAnti-Gephyrin antibodySee all Gephyrin primary antibodies ...
  • Description
    Rabbit polyclonal to Gephyrin
  • Tested applicationsICC/IF, IHC-FoFr, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat
    Predicted to work with: Chicken, Human, Xenopus laevis
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 700 to the C-terminus of Mouse Gephyrin.

    (Peptide available as ab32205.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: Brain (Mouse) Brain (Mouse) - normal tissue, 0 days old NIH 3T3 (Mouse embryonic fibroblast cell line) MEF1 (Mouse embryonic fibroblast cell line) PC12 (Rat adrenal pheochromocytoma cell line) and the following tissue lysates: Liver (Mouse) Heart (Mouse) Kidney (Mouse) Pancreas (Human) Skeletal Muscle (Mouse) Liver (Rat) Heart (Rat) Kidney (Rat)

Properties

Applications

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.

Application Abreviews Notes
ICC/IF
IHC-FoFr
WB
  • Application notesICC/IF: Use at a concentration of 1 µg/ml.
    IHC-FoFr: 1/1000.
    WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 93,75,54 kDa (predicted molecular weight: 83 kDa). Can be blocked with Gephyrin peptide (ab32205).

    NB: The 93 kDa band corresponds to Isoform 1 of Gephyrin as observed in the literature. The 54 kDa band matches the predicted size of Isoform 2. We are unsure as to the identity of the 75 kDa band but further tissue-specific isoforms of Gephyrin continue to be described and all three of the bands we observe are partially blocked by addition of ab32205.

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • FunctionMicrotubule-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.
    • PathwayCofactor biosynthesis; molybdopterin biosynthesis.
    • Involvement in diseaseDefects 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.
    • Sequence similaritiesIn the N-terminal section; belongs to the moaB/mog family.
      In the C-terminal section; belongs to the moeA family.
    • Cellular localizationCell junction > synapse. Cell junction > synapse > postsynaptic cell membrane. Cytoplasm > cytoskeleton. Cytoplasmic face of glycinergic postsynaptic membranes.
    • Information by UniProt
    • Database links
    • Alternative names
      • 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

    Anti-Gephyrin antibody images

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

      Lane 1 : Brain (Mouse) Whole Cell Lysate - Adult
      Lane 2 : Brain (Mouse) Tissue Lysate - normal tissue, 0 days old (ab7188) at 20 µg
      Lane 3 : Brain (Mouse) Whole Cell Lysate - Adult at 20 µg with Mouse Gephyrin peptide (ab32205) at 1 µg/ml
      Lane 4 : Brain (Mouse) Tissue Lysate - normal tissue, 0 days old (ab7188) at 20 µg with Mouse Gephyrin peptide (ab32205) at 1 µg/ml

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

      Performed under reducing conditions.

      Predicted band size : 83 kDa
      Observed band size : 54,75,93 kDa (why is the actual band size different from the predicted?)
    • 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. 30µm 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 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
      Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
      Lane 3 : Liver (Mouse) Tissue Lysate - normal tissue
      Lane 4 : Heart (Mouse) Tissue Lysate
      Lane 5 : Kidney (Mouse) Tissue Lysate
      Lane 6 : Pancreas (Mouse) Tissue Lysate (ab29363)
      Lane 7 : Skeletal Muscle (Mouse) Tissue Lysate (ab29711)
      Lane 8 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
      Lane 9 : Liver (Rat) Tissue Lysate
      Lane 10 : Heart (Rat) Tissue Lysate
      Lane 11 : Kidney (Rat) Whole Cell Lysate - normal tissue (ab29480)

      Lysates/proteins at 10 µg per lane.

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

      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 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab32206, 1µg/ml) for 1h 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 1h. 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 PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32206, 1µg/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 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    References for Anti-Gephyrin antibody (ab32206)

    This product has been referenced in:
    • 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) »
    • Rozic G  et al. Dynamic changes in neurexins' alternative splicing: role of Rho-associated protein kinases and relevance to memory formation. PLoS One 6:e18579 (2011). WB ; Rat . Read more (PubMed: 21533271) »

    See all 6 Publications for this product

    Product Wall

    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

    Submitted 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

    Submitted Jan 16 2008

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