Antibodies, Proteins, Kits and Reagents for Life Science

284 784 ₸ Product size

100 µl 284 784 ₸

Order now and get it on Tuesday March 02, 2021

Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR17963] to GEF H1 - C-terminal
Suitable for: ICC/IF, Flow Cyt, WB
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-GEF H1 antibody [EPR17963] - C-terminal
See all GEF H1 primary antibodies
Description

Rabbit monoclonal [EPR17963] to GEF H1 - C-terminal
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
WB	Mouse Human

See all applications and species data

Immunogen

This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa, HEK293, C6 and NIH/3T3 cell lysate. ICC/IF: HeLa and MCF-7 cells. Flow Cyt: HEK293 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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17963
Isotype

IgG
Research areas

Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

All lanes : Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ARHGEF knockout HeLa cell lysate
Lane 3 : HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 112 kDa
Observed band size: 75 kDa
why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab201687 observed at 75 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab201687 Anti-GEF H1 antibody [EPR17963] - C-terminal was shown to specifically react with GEF H1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265797 (knockout cell lysate ab257223) was used. Wild-type and GEF H1 knockout samples were subjected to SDS-PAGE. ab201687 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

All lanes : Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687) at 1/10000 dilution

Lane 1 : HEK293 (Human embryonic kidney) lysate
Lane 2 : HEK293 (Human epithelial cells from cervix adenocarcinoma) lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 112 kDa
Observed band size: 112 kDa

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

All lanes : Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lane 3 : Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) lysate
Lane 4 : C6 (Rat glial tumor cells) lysate
Lane 5 : NIH/3T3 (Mouse embyro fibroblast cells)

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 112 kDa
Observed band size: 112 kDa

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling GEF H1 with ab201687 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).
Confocal image showing cytoplasmic staining on HeLa cell line is observed.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab201687 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling GEF H1 with ab201687 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).
Confocal image showing cytoplasmic staining on MCF7 cell line is observed.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab201687 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Flow Cytometry - Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

Flow cytometric analysis of 2% paraformaldehyde-fixed HEK293 (human embryonic kidney) cells labeling GEF H1 with ab201687 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
Anti-GEF H1 antibody [EPR17963] - C-terminal (ab201687)

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