Antibodies, Proteins, Kits and Reagents for Life Science

378 595 ₸ Product size

100 µl 378 595 ₸ 10 µl 154 118 ₸

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Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR1755(2)] to APG5L/ATG5
Suitable for: ICC/IF, WB, IP, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-APG5L/ATG5 antibody [EPR1755(2)]
See all APG5L/ATG5 primary antibodies
Description

Rabbit monoclonal [EPR1755(2)] to APG5L/ATG5
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-P	Human
IP	Rat
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- Raji, HeLa, HT-1080, Human fetal kidney, C6, Raw264.7, PC-12 and NIH3T3 cell lysates; Human hepatocellular carcinoma and Human ovarian adenocarcinoma tissue
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. Stable for 12 months at -20°C.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR1755(2)
Isotype

IgG
Research areas

Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: APG5L/ATG5 knockout HAP1 cell lysate (20 µg)
Lane 3: Raji cell lysate (20 µg)
Lane 4: Jeg-3 cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab108327 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab108327 was shown to specifically react with APG5L/ATG5 when APG5L/ATG5 knockout samples were used. Wild-type and APG5L/ATG5 knockout samples were subjected to SDS-PAGE. ab108327 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Unpurified ab108327, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human ovarian adenocarcinoma tissue by Immunohistochemistry.
Immunocytochemistry/ Immunofluorescence - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Immunofluorescence staining of MCF7 cells with purified ab108327 at a working dilution of 1/150, 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 ab108327 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.
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

All lanes : Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/10000 dilution (purified)

Lane 1 : C6 cell lysate
Lane 2 : PC-12 cell lysate
Lane 3 : NIH/3T3 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 32 kDa
Observed band size: 55 kDa
why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/10000 dilution (purified) + Raji cell lysate at 20 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 32 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/2000 dilution (purified) + HT-1080 cell lysate at 20 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 32 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/2000 dilution (purified) + human fetal kidney at 10 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 32 kDa
Observed band size: 32,55 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab180327 at a working dilution of 1/150. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunoprecipitation - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

ab180327 (purified) at 1/20 immunoprecipitating CRSP8 in 10 μg PC-12 whole cell lysate (Lanes 1 and 2, observed at 55 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

All lanes : Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/1000 dilution (unpurified)

Lane 1 : Raji cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : HT-1080 cell lysate
Lane 4 : Human fetal kidney cell lysate
Lane 5 : C6 cell lysate
Lane 6 : Raw264.7 cell lysate
Lane 7 : PC-12 cell lysate
Lane 8 : NIH3T3 cell lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 32 kDa

Secondary antibody - anti-rabbit HRP (ab6721)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

Unpurified ab108327, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human hepatocellular carcinoma tissue by Immunohistochemistry.
Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327)

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