Antibodies, Proteins, Kits and Reagents for Life Science

311 587 ₸ Product size

100 µl 311 587 ₸ 10 µl 127 315 ₸

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Anti-Flotillin 1 antibody [EP446Y] (ab78178)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP446Y] to Flotillin 1
Suitable for: WB
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-Flotillin 1 antibody [EP446Y]
See all Flotillin 1 primary antibodies
Description

Rabbit monoclonal [EP446Y] to Flotillin 1
Host species

Rabbit
Tested Applications & Species

Application Species
WB
Mouse

Human

See all applications and species data
Immunogen

Synthetic peptide within Human Flotillin 1 aa 50-150. The exact sequence is proprietary.
Database link: O75955
Positive control
- WB: HEK293T, A431, HAP1, C6, K562 and HeLa cell lysates; Mouse and rat brain lysates
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
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

EP446Y
Isotype

IgG
Research areas

Western blot - Anti-Flotillin 1 antibody [EP446Y] (ab78178)

All lanes : Anti-Flotillin 1 antibody [EP446Y] (ab78178) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : FLOT1 knockout HEK293T cell lysate
Lane 3 : A431 cell lysate
Lane 4 : HAP1 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 48 kDa
Observed band size: 50 kDa
why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab78178 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab78178 Anti-Flotillin 1 antibody [EP446Y] was shown to specifically react with Flotillin 1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267276 (knockout cell lysate ab257109) was used. Wild-type and Flotillin 1 knockout samples were subjected to SDS-PAGE. ab78178 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 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Flotillin 1 antibody [EP446Y] (ab78178)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Flotillin 1 knockout HAP1 whole cell lysate (20 µg)

Lanes 1 - 2: Merged signal (red and green). Green - ab78178 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab78178 was shown to specifically react with Flotillin 1 in wild-type HAP1 cells as signal was lost in Flotillin 1 knockout cells. Wild-type and Flotillin 1 knockout samples were subjected to SDS-PAGE. ab78178 and ab9484 (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-Flotillin 1 antibody [EP446Y] (ab78178)

All lanes : Anti-Flotillin 1 antibody [EP446Y] (ab78178) at 1/2000 dilution (purified)

Lane 1 : mouse brain lysate at 10 µg
Lane 2 : rat brain lysate at 20 µg
Lane 3 : C6 cell lysate at 20 µg

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 48 kDa
Observed band size: 49 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-Flotillin 1 antibody [EP446Y] (ab78178)

All lanes : Anti-Flotillin 1 antibody [EP446Y] (ab78178) at 1/2000 dilution (purified)

Lane 1 : K562 cell lysate
Lane 2 : A431 cell lysate
Lane 3 : HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 48 kDa
Observed band size: 49 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-Flotillin 1 antibody [EP446Y] (ab78178)

Anti-Flotillin 1 antibody [EP446Y] (ab78178) at 1/500 dilution (unpurified) + K562 cell lysate

Predicted band size: 48 kDa
Observed band size: 48 kDa
Anti-Flotillin 1 antibody [EP446Y] (ab78178)

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