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    ferritin-antibody-epr3004y-ab75973.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Ferritin antibody [EPR3004Y] (ab75973)

  • Datasheet
  • SDS
Reviews (7) Submit a question References (57)

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Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)
  • Immunocytochemistry/ Immunofluorescence - Anti-Ferritin antibody [EPR3004Y] (ab75973)
  • Flow Cytometry (Intracellular) - Anti-Ferritin antibody [EPR3004Y] (ab75973)
  • Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)
  • Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)
  • Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)
  • 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 (Intra), WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

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

    Rabbit monoclonal [EPR3004Y] to Ferritin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt (Intra), WBmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • 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 (intra): 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.

Properties

  • 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

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Cardiovascular
    • Blood
    • Serum Proteins
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Signal Transduction
    • Metabolism
    • Vitamins / Minerals
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Cofactors, Vitamins / minerals
    • Vitamins / minerals
    • Metabolism
    • Types of disease
    • Neurodegenerative disease
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Anti-Ferritin antibody [EPR3004Y] - Low endotoxin, Azide free (ab215982)
    • Anti-Ferritin antibody [EPR3004Y] - BSA and Azide free (ab271854)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human FTH1 knockout HEK-293T cell line (ab266581)
  • KO cell lysates

    • Human FTH1 knockout HEK-293T cell lysate (ab256924)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab75973 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF
1/100.
Flow Cyt (Intra)
1/50.
WB (6)
1/1000 - 1/2000. Detects a band of approximately 19, 21 kDa (predicted molecular weight: 21,19 kDa).
Notes
ICC/IF
1/100.
Flow Cyt (Intra)
1/50.
WB
1/1000 - 1/2000. Detects a band of approximately 19, 21 kDa (predicted molecular weight: 21,19 kDa).
Application notes
Is unsuitable for IHC-P.

Target

  • Function

    Stores iron in a soluble, non-toxic, readily available form. Important for iron homeostasis. Has ferroxidase activity. Iron is taken up in the ferrous form and deposited as ferric hydroxides after oxidation. Also plays a role in delivery of iron to cells. Mediates iron uptake in capsule cells of the developing kidney.
  • Sequence similarities

    Belongs to the ferritin family.
    Contains 1 ferritin-like diiron domain.
  • Target information above from: UniProt accession P02794 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2495 Human
    • Entrez Gene: 2512 Human
    • Entrez Gene: 14319 Mouse
    • Entrez Gene: 14325 Mouse
    • Entrez Gene: 25319 Rat
    • Entrez Gene: 29292 Rat
    • Omim: 608847 Human
    • SwissProt: P02792 Human
    • SwissProt: P02794 Human
    • SwissProt: P09528 Mouse
    • SwissProt: P29391 Mouse
    • SwissProt: P02793 Rat
    • SwissProt: P19132 Rat
    • Unigene: 524910 Human
    • Unigene: 645560 Human
    • Unigene: 1776 Mouse
    • Unigene: 54447 Rat
    see all
  • Form

    Subunit structure: Oligomer of 24 subunits. There are two types of subunits: L (light) chain and H (heavy) chain. The major chain can be light or heavy, depending on the species and tissue type. In the human liver, the heavy chain is predominant. The functional molecule forms a roughly spherical shell with a diameter of 12 nm and contains a central cavity into which the insoluble mineral iron core is deposited.
  • Alternative names

    • Cell proliferation-inducing gene 15 protein antibody
    • Ferritin H subunit antibody
    • Ferritin heavy chain antibody
    • Ferritin heavy polypeptide 1 antibody
    • Ferritin L chain antibody
    • Ferritin L subunit antibody
    • Ferritin light chain antibody
    • Ferritin light polypeptide antibody
    • ferritin light polypeptide like 3 antibody
    • Ferritin, heavy polypeptide antibody
    • FHC antibody
    • FRIH_HUMAN antibody
    • FTH antibody
    • FTH1 antibody
    • FTHL6 antibody
    • FTL antibody
    • MGC104426 antibody
    • PIG15 antibody
    • PLIF antibody
    • Proliferation inducing protein 15 antibody
    see all

Images

  • Western blot - Anti-Ferritin antibody [EPR3004Y] (ab75973)
    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)
    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 (Intracellular) - Anti-Ferritin antibody [EPR3004Y] (ab75973)
    Flow Cytometry (Intracellular) - Anti-Ferritin antibody [EPR3004Y] (ab75973)

    Intracellular 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)
    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)
    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)
    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)
    Anti-Ferritin antibody [EPR3004Y] (ab75973)

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (57)

Publishing research using ab75973? Please let us know so that we can cite the reference in this datasheet.

ab75973 has been referenced in 57 publications.

  • Yang RZ  et al. Involvement of oxidative stress-induced annulus fibrosus cell and nucleus pulposus cell ferroptosis in intervertebral disc degeneration pathogenesis. J Cell Physiol 236:2725-2739 (2021). PubMed: 32892384
  • Jiang Z  et al. Artesunate induces ER-derived-ROS-mediated cell death by disrupting labile iron pool and iron redistribution in hepatocellular carcinoma cells. Am J Cancer Res 11:691-711 (2021). PubMed: 33791148
  • Wen J  et al. Ultrasmall iron oxide nanoparticles induced ferroptosis via Beclin1/ATG5-dependent autophagy pathway. Nano Converg 8:10 (2021). PubMed: 33796911
  • Shin HJ  et al. Lipocalin-2 Deficiency Reduces Oxidative Stress and Neuroinflammation and Results in Attenuation of Kainic Acid-Induced Hippocampal Cell Death. Antioxidants (Basel) 10:N/A (2021). PubMed: 33445746
  • Radadiya PS  et al. Ciclopirox olamine induces ferritinophagy and reduces cyst burden in polycystic kidney disease. JCI Insight 6:N/A (2021). PubMed: 33784251
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-7 of 7 Abreviews or Q&A

Immunoprecipitation abreview for Anti-Ferritin antibody [EPR3004Y]

Poor
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Mouse Cell lysate - other (primary axtrocytes)
Total protein in input
350 µg
Immuno-precipitation step
Other - protein G-Dynabeads
Specification
primary axtrocytes
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Andrea Atzmon

Verified customer

Submitted Jun 28 2022

Western blot abreview for Anti-Ferritin antibody [EPR3004Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (SY5Y cell)
Gel Running Conditions
Reduced Denaturing (4-20% gel)
Loading amount
15 µg
Specification
SY5Y cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 12 2021

Western blot abreview for Anti-Ferritin antibody [EPR3004Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (RAW264.7)
Gel Running Conditions
Reduced Denaturing (15% gel)
Loading amount
50000 cells
Specification
RAW264.7
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 27 2020

Western blot abreview for Anti-Ferritin antibody [EPR3004Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Liver - Hepg2)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
30 µg
Treatment
50uM Ferric Citrate
Specification
Liver - Hepg2
Blocking step
Licor Odyssey blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Boyd Kenkhuis

Verified customer

Submitted May 06 2020

Western blot abreview for Anti-Ferritin antibody [EPR3004Y]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Zebrafish Tissue lysate - whole (Eye, muscle)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris gel, NuPAGE)
Loading amount
23 µg
Specification
Eye, muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Cheri Ackerman

Verified customer

Submitted Aug 11 2016

Western blot abreview for Anti-Ferritin antibody [EPR3004Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (TGX any size gel)
Sample
Fish Cell lysate - whole cell (Liver)
Specification
Liver
Blocking step
(agent) for 20 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Don Kuhn

Verified customer

Submitted Oct 29 2013

Western blot abreview for Anti-Ferritin antibody [EPR3004Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Loading amount
20 µg
Specification
liver
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 06 2011

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