AB325248

Anti-Beclin-1 (phospho S90) antibody [EPR26903-61R] - BSA and Azide free

RabMAb
Recombinant
KO Validated
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Rabbit Recombinant Monoclonal Beclin 1 phospho S90 antibody. Carrier free. Suitable for WB, Dot and reacts with Transfected cell lysate - Human, Human, Synthetic peptide - Human samples.

View Alternative Names

GT197, Beclin-1, Coiled-coil myosin-like BCL2-interacting protein, Protein GT197

4 Images

Supplier Data

Western blot - Anti-Beclin-1 (phospho S90) antibody [EPR26903-61R] - BSA and Azide free (AB325248)

This data was developed using ab325242, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab325242 was shown to react with Beclin-1 (phospho S90) in wild-type HeLa serum starved overnight then treated with 100nM Calyculin A for 30 minutes lysate(lane2) with loss of signal observed in BECN1 knockout HeLa serum starved overnight then treated with 100nM Calyculin A for 30 minutes lysate(lane3).

The identity of the higher MW bands at approximately 70KDa and 250KDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Beclin 1 antibody [EPR19662] (ab207612) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Beclin-1 (phospho S90) antibody [EPR26903-61R] (<a href='/en-us/products/primary-antibodies/beclin-1-phospho-s90-antibody-epr26903-61r-ab325242'>ab325242</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 80 µg

Lane 2:

HeLa serum starved overnight then treated with 100nM Calyculin A for 30 minutes, whole cell lysate (untreated membrane) at 80 µg

Lane 3:

BECN1 knock-out HeLa serum starved overnight then treated with 100nM Calyculin A for 30 minutes, whole cell lysate (untreated membrane) at 80 µg

Lane 4:

HeLa serum starved overnight then treated with 100nM Calyculin A for 30 minutes, whole cell lysate (phosphatase treated membrane) at 80 µg

Lane 5:

BECN1 knock-out HeLa serum starved overnight then treated with 100nM Calyculin A for 30 minutes, whole cell lysate (phosphatase treated membrane) at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 60 kDa,36 kDa,52 kDa

false

Exposure time: 180s

Supplier Data

Western blot - Anti-Beclin-1 (phospho S90) antibody [EPR26903-61R] - BSA and Azide free (AB325248)

This data was developed using ab325242, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the higher MW band at approximately 70KDa is unknown

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Beclin 1 antibody [EPR19662] (ab207612) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Beclin-1 (phospho S90) antibody [EPR26903-61R] (<a href='/en-us/products/primary-antibodies/beclin-1-phospho-s90-antibody-epr26903-61r-ab325242'>ab325242</a>) at 1/1000 dilution

Lane 1:

Untreated HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 80 µg

Lane 2:

HEK-293 treated with 100nM Calyculin A for 30 minutes whole cell lysate at 80 µg

Lane 3:

Untreated MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 80 µg

Lane 4:

MCF7 treated with 100nM Calyculin A for 30 minutes whole cell lysate at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 60 kDa,36 kDa,52 kDa

false

Exposure time: 180s

Supplier Data

Western blot - Anti-Beclin-1 (phospho S90) antibody [EPR26903-61R] - BSA and Azide free (AB325248)

This data was developed using ab325242, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

CaMKII has been shown to directly phosphorylate Beclin 1 at Ser90 (PMID : 29079782)

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

In Western blot, Anti-Beclin 1 antibody [EPR19662] (ab207612) staining at 1/1000 dilution.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Beclin-1 (phospho S90) antibody [EPR26903-61R] (<a href='/en-us/products/primary-antibodies/beclin-1-phospho-s90-antibody-epr26903-61r-ab325242'>ab325242</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T transfected with human BECN1 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with human BECN1 expression vector containing a myc-His-tag® transfected with a human wild-type CaMKII beta expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 4:

293T transfected with human BECN1(S90A mutant) expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60 kDa,80 kDa,52 kDa,36 kDa

false

Exposure time: 8s

Supplier Data

Dot Blot - Anti-Beclin-1 (phospho S90) antibody [EPR26903-61R] - BSA and Azide free (AB325248)

This data was developed using ab325242, the same antibody clone in a different buffer formulation.

Dot blot analysis of Phospho-Beclin-1 (Ser90) using ab325242 at 1 : 1000 (0.471 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Human Beclin-1 (phospho S90) peptide
Lane2 : Human Beclin-1 (non-phospho) peptide
Lane3 : Human Beclin-1 (phospho S96) peptide
Lane4 : Human Beclin-1 (phospho S93) peptide

Exposure time : 180 seconds.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with Human Beclin-1 (phospho S93) or (phospho S96) peptides by dot blot.

All lanes:

Dot Blot - Anti-Beclin-1 (phospho S90) antibody [EPR26903-61R] (<a href='/en-us/products/primary-antibodies/beclin-1-phospho-s90-antibody-epr26903-61r-ab325242'>ab325242</a>) at 1/1000 dilution

Lane 1:

Human Beclin-1 (phospho S90) peptide

Lane 2:

Human Beclin-1 (non-phospho) peptide

Lane 3:

Human Beclin-1 (phospho S96) peptide

Lane 4:

Human Beclin-1 (phospho S93) peptide

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR26903-61R

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB, Dot

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

We have observed a non-specific band at ~70KDa using calyculin A to induce phosphorylation. We suggest to include total Beclin-1 detection as a control to confirm Beclin-1 molecular weight (~60KDa)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "Synthetic peptide - Human": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "Transfected cell lysate - Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }

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Product details

ab325248 is the carrier-free version of ab325242.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

Plays a central role in autophagy (PubMed : 18570871, PubMed : 21358617, PubMed : 23184933, PubMed : 23974797, PubMed : 25484083, PubMed : 28445460, PubMed : 37776275). Acts as a core subunit of the PI3K complex that mediates formation of phosphatidylinositol 3-phosphate; different complex forms are believed to play a role in multiple membrane trafficking pathways : PI3KC3-C1 is involved in initiation of autophagosomes and PI3KC3-C2 in maturation of autophagosomes and endocytosis. Involved in regulation of degradative endocytic trafficking and required for the abscission step in cytokinesis, probably in the context of PI3KC3-C2 (PubMed : 20208530, PubMed : 20643123, PubMed : 23974797, PubMed : 26783301). Essential for the formation of PI3KC3-C2 but not PI3KC3-C1 PI3K complex forms. Involved in endocytosis (PubMed : 25275521). May play a role in antiviral host defense.. Beclin-1-C 35 kDa localized to mitochondria can promote apoptosis; it induces the mitochondrial translocation of BAX and the release of proapoptotic factors.. (Microbial infection) Protects against infection by a neurovirulent strain of Sindbis virus.

See full target information BECN1 phospho S90

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Product promise

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For full details, please see our Terms & Conditions

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