This antibody gave a positive signal in the following whole cell lysates:
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:
Skeletal Muscle (Mouse)
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
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).
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.
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.
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.
Western blot - Gephyrin antibody (ab32206)
All lanes : Anti-Gephyrin antibody (ab32206) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse) 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)
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 A110:14777-82 (2013).
Read more (PubMed: 23959889) »
Hales CM et al. Abnormal gephyrin immunoreactivity associated with Alzheimer disease pathologic changes. J Neuropathol Exp Neurol72:1009-15 (2013).
Read more (PubMed: 24128675) »