Antibodies, Proteins, Kits and Reagents for Life Science

345 091 ₸ Product size

100 µl 345 091 ₸

Order now and get it on Tuesday March 02, 2021

Anti-Ferritin antibody [EPR3004Y] (ab75973)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR3004Y] to Ferritin
Suitable for: ICC/IF, Flow Cyt, WB
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-Ferritin antibody [EPR3004Y]
See all Ferritin primary antibodies
Description

Rabbit monoclonal [EPR3004Y] to Ferritin
Host species

Rabbit
Specificity

In human, this antibody recognizes both, Ferritin heavy and light chains. In mouse and rat, it only recognizes Ferritin heavy chain.

Tested Applications & Species

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

See all applications and species data

Immunogen

Synthetic peptide within Human Ferritin aa 50-150. The exact sequence is proprietary.
Database link: P02794
Positive control
- WB: HEK293T, Jurkat, SH-SY-5Y, C6 and Raw264.7 cell lysates; Human and rat brain lysates ICC/IF: Jurkat cells Flow Cyt: SH-SY5Y 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.

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: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR3004Y
Isotype

IgG
Research areas

Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)

All lanes : Anti-Ferritin antibody [EPR3004Y] (ab75973) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : FTH1 knockout HEK293T cell lysate
Lane 3 : SH-SY5Y 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: 21,19 kDa
Observed band size: 21 kDa
why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab75973 observed at 21 kDa. Red - loading control ab8245 observed at 36 kDa.

ab75973 Anti-Ferritin antibody [EPR3004Y] was shown to specifically react with Ferritin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266581 (knockout cell lysate ab256924) was used. Wild-type and Ferritin knockout samples were subjected to SDS-PAGE. ab75973 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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.
Immunocytochemistry/ Immunofluorescence - Anti-Ferritin antibody [EPR3004Y] (ab75973)

Immunofluorescence staining of Jurkat cells with purified ab75973 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab75973 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Flow Cytometry - Anti-Ferritin antibody [EPR3004Y] (ab75973)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling Ferritin with ab75973 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)

All lanes : Anti-Ferritin antibody [EPR3004Y] (ab75973) at 1/2000 dilution

Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 3 : 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), Peroxidase conjugated at 1/20000 dilution

Predicted band size: 21,19 kDa

Observed MW: 21 kDA

Blocking/diluting buffer and concentration: 5% NFDM/TBST
Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)

Anti-Ferritin antibody [EPR3004Y] (ab75973) at 1/10000 dilution (purified) + Raw264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 21,19 kDa
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-Ferritin antibody [EPR3004Y] (ab75973)

All lanes : Anti-Ferritin antibody [EPR3004Y] (ab75973) at 1/2000 dilution (Unpurified)

Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate
Lane 2 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) cell lysate
Lane 3 : Human brain lysates

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 21,19 kDa
Observed band size: 19,21 kDa why is the actual band size different from the predicted?
Anti-Ferritin antibody [EPR3004Y] (ab75973)

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