Antibodies, Proteins, Kits and Reagents for Life Science

311 587 ₸ Product size

100 µl 311 587 ₸

Order now and get it on Tuesday March 09, 2021

Anti-Gephyrin antibody [EPR12651(B)] (ab177154)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR12651(B)] to Gephyrin
Suitable for: ICC, WB, IHC-P, Flow Cyt
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-Gephyrin antibody [EPR12651(B)]
See all Gephyrin primary antibodies
Description

Rabbit monoclonal [EPR12651(B)] to Gephyrin
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC	Rat Human
IHC-P	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Gephyrin aa 700 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
Database link: Q9NQX3
Positive control
- WB: Human fetal brain, 293T, Jurkat, SH-SY5Y, C6, Raw264.7, MCF-7, U2OS, and PC-12 lysates; IHC-P: Human brain tissue; Flow Cyt: Permeabilized 293T cells; ICC: Jurkat and PC-12 cells.
General notes
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR12651(B)
Isotype

IgG
Research areas

Immunocytochemistry - Anti-Gephyrin antibody [EPR12651(B)] (ab177154)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-12 cells labelling Gephyrin with ab177154 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing cytoplasmic staining in PC-12 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: PBS instead of the primary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Gephyrin antibody [EPR12651(B)] (ab177154)

Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling Gephyrin with ab177154 at 1/50 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Western blot - Anti-Gephyrin antibody [EPR12651(B)] (ab177154)

All lanes : Anti-Gephyrin antibody [EPR12651(B)] (ab177154) at 1 µg/ml

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

Lanes 1 - 4: Merged signal (red and green). Green - ab177154 observed at 90 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab177154 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. Ab177154 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.
Flow Cytometry - Anti-Gephyrin antibody [EPR12651(B)] (ab177154)

Flow cytometric analysis of permeabilized 293T cells labeling Gephyrin with ab177154 at 1/10 dilution (red), or a rabbit IgG (negative) (green).
Immunocytochemistry - Anti-Gephyrin antibody [EPR12651(B)] (ab177154)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat cells labelling Gephyrin with ab177154 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing cytoplasmic staining in Jurkat cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: PBS instead of the primary antibody.
Western blot - Anti-Gephyrin antibody [EPR12651(B)] (ab177154)

All lanes : Anti-Gephyrin antibody [EPR12651(B)] (ab177154) at 1/1000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : 293T lysate
Lane 3 : Jurkat lysate
Lane 4 : SH-SY5Y lysate
Lane 5 : C6 lysate
Lane 6 : Raw264.7 lysate
Lane 7 : PC-12 lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 80 kDa
Anti-Gephyrin antibody [EPR12651(B)] (ab177154)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com