Overview

  • Product name
  • Description
    Rabbit polyclonal to Gephyrin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat
    Predicted to work with: Chicken, Human, Xenopus laevis
  • Immunogen

    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)

  • Positive control
    • WB: Mouse brain and liver tissue lysate, rat brain and liver tissue lysate. ICC/IF: PC-12 cells.

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 Use a concentration of 1 µg/ml.
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).

Target

  • Function
    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.
  • Pathway
    Cofactor biosynthesis; molybdopterin biosynthesis.
  • Involvement in disease
    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.
  • Sequence similarities
    In the N-terminal section; belongs to the moaB/mog family.
    In the C-terminal section; belongs to the moeA family.
  • Cellular localization
    Cell 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

Images

  • All lanes : ab32206 at 1 µg/ml

    Lane 1 : Mouse Brain Tissue Lysate
    Lane 2 : Mouse Liver Tissue Lysate
    Lane 3 : Rat Brain Tissue Lysate
    Lane 4 : Rat Liver Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/1000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 83 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 minutes


    We are unsure of the identity of the 89kDa band observed in brain lysates, however this was consistent across both species tested. 

    Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

  • 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.

  • 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.

References

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) »
  • Notaras M  et al. The BDNF Val66Met polymorphism regulates glucocorticoid-induced corticohippocampal remodeling and behavioral despair. Transl Psychiatry 7:e1233 (2017). WB ; Mouse . Read more (PubMed: 28926000) »
See all 12 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or 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

Abcam user community

Verified customer

Submitted 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

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

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"

Sign up