Antibodies, Proteins, Kits and Reagents for Life Science

294 835 ₸ Product size

100 µl 294 835 ₸

Order now and get it on Tuesday March 02, 2021

Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20451] to SQSTM1 / p62 (phospho S349)
Suitable for: ICC/IF, IP, Flow Cyt, WB
Reacts with: Mouse, Rat, Human

Product name

Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451]
See all SQSTM1 / p62 primary antibodies
Description

Rabbit monoclonal [EPR20451] to SQSTM1 / p62 (phospho S349)
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IP	Human
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
- WB: HeLa, C6 and NIH/3T3 whole cell lysates treated with 2µM MG-132 (ab141003) for 18 hours. ICC/IF: HeLa cells treated with 2µM MG-132 (ab141003) for 18 hours. Flow Cyt: HeLa cells treated with 2µM MG-132 (ab141003) for 18 hours. IP: HeLa treated with 2µM MG-132 (ab141003) for 18h whole cell lysate.
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.

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR20451
Isotype

IgG
Research areas

Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution

Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours
Lane 3 : HeLa whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours, then treated with Alkaline Phosphatase for 1 hour

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 47 kDa
Observed band size: 62 kDa
why is the actual band size different from the predicted?

Exposure time: 1 minute

Blocking/dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 2μM MG-132 (ab141003) for 18 hours or untreated, labeling SQSTM1 / p62 (phospho S349) with ab211324 at 1/100 dilution, followed by Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) secondary antibody at 1/200 dilution (green).

Confocal image showing cytoplasmic staining on HeLa cell line. The expression increased after treatment with 2μM MG-132 (ab141003) for 18 hours.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Flow Cytometry - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 2μM MG-132 (ab141003) for 18 hours (red) or untreated (green), labeling SQSTM1 / p62 (phospho S349) with ab211324 at 1/500 dilution compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

SQSTM1 / p62 (phospho S349) was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 2μM MG-132 (ab141003) for 18h with ab211324 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab211324 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1: HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate, 10 µg (Input).
Lane 2: ab211324 IP in HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211324 in HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate.

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution

Lane 1 : Untreated C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : C6 whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 47 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution

Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 47 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?

Exposure time: 4 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

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