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) 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.

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

Predicted band size: 20 kDa

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.

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.

 

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.

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

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

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