Anti-Ferritin Light Chain antibody [FTL/1386] KO Tested (ab218400)

Product name

Anti-Ferritin Light Chain antibody [FTL/1386]
See all Ferritin Light Chain primary antibodies
Description

Mouse monoclonal [FTL/1386] to Ferritin Light Chain
Host species

Mouse
Tested applications

Suitable for: Protein Array, WB, IHC-Pmore details
Species reactivity

Reacts with: Human
Immunogen

Recombinant fragment within Human Ferritin Light Chain aa 38-165. The exact sequence is proprietary.
Database link: P02792
Positive control
- WB: HAP1, A549, A431, HeLa cell lysates; Human liver, kidney, and testis lysates; IHC-P: Human pancreas and testicular carcinoma tissues.
General notes

The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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: 0.05% Sodium azide
Constituents: 99% PBS, 0.05% BSA
Concentration information loading...
Purity

Protein A/G purified
Purification notes

ab218400 is purified from Bioreactor Concentrate by Protein A/G.
Clonality

Monoclonal
Clone number

FTL/1386
Isotype

IgG2b
Research areas

Alternative Versions
- Anti-Ferritin Light Chain antibody [FTL/1386] - BSA and Azide free (ab218866)
Isotype control
- Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
KO cell lines
- Human FTL knockout HeLa cell line (ab265533)
- Human FTL knockout HeLa cell line (ab265534)
KO cell lysates
- Human FTL knockout HeLa cell lysate (ab256926)
- Human FTL knockout HeLa cell lysate (ab256927)
Positive Controls
Recombinant Protein
- Recombinant Human Ferritin Light Chain protein (ab158473)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab218400 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
Protein Array		Use at an assay dependent concentration.
WB		Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 20 kDa.
IHC-P		Use a concentration of 0.25 - 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Notes
Protein Array Use at an assay dependent concentration.
WB Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 20 kDa.
IHC-P Use a concentration of 0.25 - 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Function

Stores iron in a soluble, non-toxic, readily available form. Important for iron homeostasis. 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.
Involvement in disease

Defects in FTL are the cause of hereditary hyperferritinemia-cataract syndrome (HHCS) [MIM:600886]. It is an autosomal dominant disease characterized by early-onset bilateral cataract. Affected patients have elevated level of circulating ferritin. HHCS is caused by mutations in the iron responsive element (IRE) of the FTL gene.
Defects in FTL are the cause of neurodegeneration with brain iron accumulation type 3 (NBIA3) [MIM:606159]; also known as adult-onset basal ganglia disease. It is a movement disorder with heterogeneous presentations starting in the fourth to sixth decade. It is characterized by a variety of neurological signs including parkinsonism, ataxia, corticospinal signs, mild nonprogressive cognitive deficit and episodic psychosis. It is linked with decreased serum ferritin levels.
Sequence similarities

Belongs to the ferritin family.
Contains 1 ferritin-like diiron domain.
Target information above from: UniProt accession P02792 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 2512 Human
- Omim: 134790 Human
- SwissProt: P02792 Human
- Unigene: 433670 Human
- Unigene: 713706 Human
Alternative names
- Ferritin L chain antibody
- Ferritin L subunit antibody
- Ferritin light chain antibody
- Ferritin light polypeptide antibody
- ferritin light polypeptide like 3 antibody
- FRIL_HUMAN antibody
- FTL antibody
- LFTD antibody
- NBIA 3 antibody
- NBIA3 antibody
see all

Western blot - Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400)

All lanes : Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : FTL knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 20 kDa
Observed band size: 20 kDa

Lanes 1- 2: Merged signal (red and green). Green - ab218400 observed at 20 kDa. Red - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) observed at 50 kDa.

ab218400 was shown to react with Ferritin Light Chain in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265534 (knockout cell lysate ab256927) was used. Wild-type HeLa and FTL knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab218400 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testicular carcinoma tissue labelling Ferritin Light Chain with ab218400 at 0.5 µg/mL (30 minutes at room temperature). Staining of formalin-fixed tissues requires boiling tissue sections in 10mM Citrate Buffer, pH 6.0, for 10-20 minutes followed by cooling at room temperature for 20 minutes.
Western blot - Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400)

Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400) at 2 µg/ml + Human kidney tissue lysate

Predicted band size: 20 kDa
Protein Array - Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400)

ab218400 was tested in protein array against over 19000 different full-length human proteins.
Z- and S- Score: The Z-score represents the strength of a signal that a monoclonal antibody (MAb) (in combination with a fluorescently-tagged anti-IgG secondary antibody) produces when binding to a particular protein on the HuProtTM array. Z-scores are described in units of standard deviations (SD's) above the mean value of all signals generated on that array. If targets on HuProtTM are arranged in descending order of the Z-score, the S-score is the difference (also in units of SD's) between the Z-score. S-score therefore represents the relative target specificity of a MAb to its intended target.
A MAb is specific to its intended target if the MAb has an S-score of at least 2.5. For example, if a MAb binds to protein X with a Z-score of 43 and to protein Y with a Z-score of 14, then the S-score for the binding of that MAb to protein X is equal to 29.
Western blot - Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400)

All lanes : Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : FTL knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 20 kDa

Lanes 1-2: Merged signal (red and green). Green - ab218400 observed at 20 kDa. Red - loading control ab181602 observed at 37 kDa.

ab218400 Anti-FTL was shown to specifically react with Ferritin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265533 (knockout cell lysate ab256926) was used. Wild-type and FTL knockout samples were subjected to SDS-PAGE. ab218400 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400)

All lanes : Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : FTL (Ferritin Light Chain) knockout HAP1 whole cell lysate
Lane 3 : A549 whole cell lysate
Lane 4 : Hep G2 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 20 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab218400 observed at 20 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab218400 was shown to recognize Ferritin Light Chain in wild-type HAP1 cells as signal was lost at the expected MW in FTL (Ferritin Light Chain) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and FTL (Ferritin Light Chain) knockout samples were subjected to SDS-PAGE. Ab218400 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400)

All lanes : Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400) at 0.2 µg/ml

Lanes 1-2 : HeLa cell lysate
Lane 3 : Human liver lysate
Lane 4 : Human testis lysate

Predicted band size: 20 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human pancreas tissue labeling Ferritin Light Chain with ab218400 at 0.2 μg/ml.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ferritin Light Chain antibody [FTL/1386] (ab218400)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human pancreas tissue labeling Ferritin Light Chain with ab218400 at 0.2 μg/ml.

Western blot protocols
Immunohistochemistry protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab218400? Please let us know so that we can cite the reference in this datasheet.

ab218400 has been referenced in 2 publications.

Terzi EM et al. Iron-sulfur cluster deficiency can be sensed by IRP2 and regulates iron homeostasis and sensitivity to ferroptosis independent of IRP1 and FBXL5. Sci Adv 7:N/A (2021). PubMed: 34039609
Chen X et al. Andrographolide Suppresses MV4-11 Cell Proliferation through the Inhibition of FLT3 Signaling, Fatty Acid Synthesis and Cellular Iron Uptake. Molecules 22:N/A (2017). PubMed: 28858244

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Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Purification notes

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

References (2)

Customer reviews and Q&As