Anti-ATG4A antibody (ab223374)
Key features and details
- Rabbit polyclonal to ATG4A
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-ATG4A antibody
See all ATG4A primary antibodies -
Description
Rabbit polyclonal to ATG4A -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB RatHuman
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Immunogen
Synthetic peptide corresponding to Human ATG4A aa 100-200. From the mid-protein.
Database link: Q8WYN0 -
Positive control
- WB: HeLa and Daudi cell lysates; Rat pancreas cell lysate. ICC/IF: SK-N-BE cells.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
Properties
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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
Preservative: 0.09% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-ATG4A antibody (ab223374)All lanes : Anti-ATG4A antibody (ab223374) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ATG4A knockout HeLa cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 37,39, 42, 45 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab223374 observed at 45 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab223374 was shown to react with ATG4A in wild-type cells in Western blot with loss of signal observed in ATG4A knockout cell line ab265738 (ATG4A knockout cell lysate ab257846). Wild-type and ATG4A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab223374 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-ATG4A antibody (ab223374)4% formaldehyde-fixed SK-N-BE cells stained for ATG4A (green) using ab223374 at 1/100 dilution in ICC/IF. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1/1000, 1/5000 for 60 minutes RT, 5 minutes RT.
(A) DAPI nuclear stain (B) Phalloidin Texas Red F-Actin stain (C) ab223374 (D) Merge.
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Western blot - Anti-ATG4A antibody (ab223374)Anti-ATG4A antibody (ab223374) at 1/1000 dilution + rat pancreas cell lysate at 15 µg
Secondary
Goat Anti-Rabbit HRP at 1/2000 dilution
Developed using the ECL technique.
Predicted band size: 37,39, 42, 45 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 6 minutesBlocking buffer: 5% skimmed milk in TBST.
Primary and secondary antibodies were incubated for 1 hour at RT.
