Recombinant Anti-Gephyrin antibody [EPR12650] - BSA and Azide free (ab250503)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12650] to Gephyrin - BSA and Azide free
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Gephyrin antibody [EPR12650] - BSA and Azide free
See all Gephyrin primary antibodies -
Description
Rabbit monoclonal [EPR12650] to Gephyrin - BSA and Azide free -
Host species
Rabbit -
Specificity
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Cow, Dog, Pig, Zebrafish, Rhesus monkey, Xenopus tropicalis -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293, Mouse heart, Fetal brain, SH-SY5Y, Neuro-2a, HAP1, MCF7, U2OS, and C6 lysates; IHC-P: Human kidney and brain tissue.
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General notes
ab250503 is the carrier-free version of ab181382.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR12650 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab250503 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 |
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WB |
1/1000 - 1/5000. Detects a band of approximately 93 kDa (predicted molecular weight: 80 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
Notes |
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WB
1/1000 - 1/5000. Detects a band of approximately 93 kDa (predicted molecular weight: 80 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. |
Target
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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
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Database links
- Entrez Gene: 428878 Chicken
- Entrez Gene: 535194 Cow
- Entrez Gene: 490739 Dog
- Entrez Gene: 10243 Human
- Entrez Gene: 268566 Mouse
- Entrez Gene: 100153443 Pig
- Entrez Gene: 64845 Rat
- Entrez Gene: 709677 Rhesus monkey
see all -
Alternative names
- Domain E antibody
- Domain G antibody
- GEPH antibody
see all
Images
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This data was developed using ab181382, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of parafiin embedded human brain tissue using ab181382 (unpurified).
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemical analysis of parafiin embedded human brain tissue using ab181382 (unpurified).
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-Gephyrin antibody [EPR12650] (ab181382) at 1/1000 dilution (Unpurified)
Lane 1 : Fetal brain tissue lysate
Lane 2 : 293T cell lysate
Lane 3 : SH-SY5Y cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 80 kDaThis data was developed using ab181382, the same antibody clone in a different buffer formulation.
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All lanes : Anti-Gephyrin antibody [EPR12650] (ab181382) at 1/1000 dilution (purified)
Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 2 : Mouse heart lysates
Lane 3 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates
Lane 4 : C6 (Rat glial tumor glial cell) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 80 kDa
Observed band size: 93 kDa why is the actual band size different from the predicted?This data was developed using ab181382, the same antibody clone in a different buffer formulation.
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All lanes : Anti-Gephyrin antibody [EPR12650] (ab181382) at 1 µg/ml (unpurified)
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : GPHN (Gephyrin) knockout HAP1 whole cell lysate
Lane 3 : MCF7 whole cell lysate
Lane 4 : U2OS whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 80 kDa
This data was developed using ab181382, the same antibody clone in a different buffer formulation.
Lanes 1 - 4: Merged signal (red and green). Green - ab181382 observed at 90 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab181382 was shown to recognize Gephyrin in wild-type HAP1 cells as signal was lost at the expected MW in GPHN (Gephyrin) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GPHN (Gephyrin) knockout samples were subjected to SDS-PAGE. Ab181382 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling Gephyrin with purified ab181382 at 1/100 dilution (1.62 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181382)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab250503 has not yet been referenced specifically in any publications.