Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 09, 2021

Anti-GTPase HRAS antibody [Y132] (ab32417)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y132] to GTPase HRAS
Suitable for: WB, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-GTPase HRAS antibody [Y132]
See all GTPase HRAS primary antibodies
Description

Rabbit monoclonal [Y132] to GTPase HRAS
Host species

Rabbit
Specificity

Reactivity with other RAS members has not been tested.
Tested Applications & Species

Application Species
IP
Mouse

WB
Mouse

Human

See all applications and species data
Immunogen

Synthetic peptide within Human GTPase HRAS aa 150 to the C-terminus (C terminal). The exact sequence is proprietary.
Positive control
- MCF7 and PC12 cell lysates and MCF7 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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Application	Species
IP	Mouse
WB	Mouse Human

Form

Liquid
Storage instructions

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

pH: 7.20
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

Y132
Isotype

IgG
Research areas

Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)

All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/500 dilution

Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : HRAS knockout HEK-293 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate

Lysates/proteins at 20 µg/ml per lane.

Lanes 1 - 4: Merged signal (red and green). Green - ab32417 observed at 21 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab32417 was shown to recognize HRAS in wild-type HEK-293 cells as signal was lost at the expected MW in HRAS knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and HRAS knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab32417 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution 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.
Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)

All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/2500 dilution (purified)

Lane 1 : mouse brain lysate
Lane 2 : rat brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

Observed band size: 21 kDa
why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)

All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/1000 dilution (purified)

Lane 1 : MCF7 cell lysate
Lane 2 : HeLa cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

Observed band size: 21 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunoprecipitation - Anti-GTPase HRAS antibody [Y132] (ab32417)

ab32417 (purified) at 1/60 immunoprecipitating GTPase in 10 μg mouse brain whole cell lysate (Lanes 1 and 2, observed at 21 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP Detection Reagent (ab131366) was used for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)

All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/500 dilution (unpurified)

Lane 1 : MCF7 cell lysate
Lane 2 : PC12 cell lysate

Observed band size: 21 kDa why is the actual band size different from the predicted?
Anti-GTPase HRAS antibody [Y132] (ab32417)

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