AB303550

Anti-Bad (phospho S99) antibody [EPR24751-161]

20ul selling size
RabMAb
Recombinant
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Rabbit Recombinant Monoclonal BAD phospho S99 antibody. Suitable for WB, Dot and reacts with Human, Transfected cell line - Human, Synthetic peptide - Human samples.

View Alternative Names

BBC6, BCL2L8, BAD, Bcl2-associated agonist of cell death, Bcl-2-binding component 6, Bcl-2-like protein 8, Bcl-xL/Bcl-2-associated death promoter, Bcl2 antagonist of cell death, Bcl2-L-8

3 Images

Lab

Western blot - Anti-Bad (phospho S99) antibody [EPR24751-161] (AB303550)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining.

All lanes:

Western blot - Anti-Bad (phospho S99) antibody [EPR24751-161] (ab303550) at 1/1000 dilution

Lane 1:

Untreated HT-29 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

HT-29 cells treated with 0.1uM Calyculin for 25 min, 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 1/20000 dilution

Observed band size: 20-37 kDa

false

Exposure time: 180s

Lab

Western blot - Anti-Bad (phospho S99) antibody [EPR24751-161] (AB303550)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining.

All lanes:

Western blot - Anti-Bad (phospho S99) antibody [EPR24751-161] (ab303550) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney epithelial cell) transfected with a human BAD expression vector containing a his tag was treated with 0.1uM Calyculin for 25 min, whole cell lysate (untreated membrane) at 20 µg

Lane 2:

HEK-293T cells transfected with a human BAD expression vector containing a his tag was treated with 0.1uM Calyculin for 25 min, whole cell lysate (phosphatase treated membrane) 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: 20-37 kDa

false

Exposure time: 15s

Lab

Dot Blot - Anti-Bad (phospho S99) antibody [EPR24751-161] (AB303550)

Dot blot analysis of Bad using ab303550 at 1/1000 followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.

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

All lanes:

Dot Blot - Anti-Bad (phospho S99) antibody [EPR24751-161] (ab303550) at 1/1000 dilution

Lane 1:

BAD phospho S99 peptide

Lane 2:

BAD non-phospho peptide corresponding to the region covered by BAD pS99 -containing 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

Carrier free

Anti-Bad (phospho S99) antibody [EPR24751-161] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR24751-161

Isotype

IgG

Carrier free

Reacts with

Human

Applications

WB, Dot

applications

Immunogen

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

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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"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "Observed Mass of target (kDa) 20-37,23", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Mouse": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Rat": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Synthetic peptide - Human": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Transfected cell line - Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "Observed Mass of target (kDa) 20-37,23", "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" } } }

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Product details

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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The Bad protein also known as Bcl-2-associated death promoter plays a critical role in apoptosis regulation. It possesses a molecular weight of approximately 23 kDa. Bad is expressed widely in tissues and is a member of the Bcl-2 family of proteins. This family is well-known for its involvement in the regulation of cell death both promoting and inhibiting depending on the protein’s interactions.

Biological function summary

Its main role is to promote apoptosis by binding to anti-apoptotic proteins like Bcl-2 and Bcl-XL displacing pro-apoptotic Bax and Bak proteins to trigger cell death. Bad functions as part of the mitochondrial apoptosis pathway and is known to form complexes with phosphorylated proteins. Phosphorylation of Bad by kinases can lead to its sequestration to the cytoplasm reducing apoptosis.

Pathways

Its action is pivotal in the PI3K/Akt signaling pathway. This pathway involves growth factors and cell survival signals where phosphorylation of Bad inhibits its pro-apoptotic activity. Bad interacts with proteins like Akt and 14-3-3 to modulate cell survival. Disruption in its regulation through these pathways may lead to abnormal cell survival or death.

Bad has implications in cancer and neurodegenerative diseases. Cancer cells often exhibit impaired apoptotic pathways where overexpression of Bad can ameliorate responsiveness to therapy. In neurodegenerative diseases such as Alzheimer's dysregulation of its function can contribute to cell death. The protein’s interaction with Bcl-2 and Bcl-XL is significant in these contexts influencing disease propagation and severity.

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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

Promotes cell death. Successfully competes for the binding to Bcl-X(L), Bcl-2 and Bcl-W, thereby affecting the level of heterodimerization of these proteins with BAX. Can reverse the death repressor activity of Bcl-X(L), but not that of Bcl-2 (By similarity). Appears to act as a link between growth factor receptor signaling and the apoptotic pathways.

See full target information BAD phospho S99

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-Bad (phospho S99) antibody [EPR24751-161]

Western blot - Anti-Bad (phospho S99) antibody [EPR24751-161] (AB303550)

Western blot - Anti-Bad (phospho S99) antibody [EPR24751-161] (AB303550)

Dot Blot - Anti-Bad (phospho S99) antibody [EPR24751-161] (AB303550)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Complementary Products

Primary Antibodies

Primary Antibodies

Secondary Antibodies

Primary Antibodies

Product promise