AB245824

Anti-Flt3 / CD135 antibody [EPR22232-318] - BSA and Azide free

RabMAb
Recombinant
KO Validated
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Knockout Tested Rabbit Recombinant Monoclonal Flt3 / CD135 antibody. Carrier free. Suitable for IP, WB and reacts with Human samples.

View Alternative Names

CD135, FLK2, STK1, FLT3, Receptor-type tyrosine-protein kinase FLT3, FL cytokine receptor, Fetal liver kinase-2, Fms-like tyrosine kinase 3, Stem cell tyrosine kinase 1, FLK-2, FLT-3, STK-1

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Immunoprecipitation - Anti-Flt3 / CD135 antibody [EPR22232-318] - BSA and Azide free (AB245824)

Flt3 / CD135 was immunoprecipitated from 0.35 mg of THP-1 (human monocytic leukemia cell line) whole cell lysate with ab245116 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab245116 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1 : THP-1 whole cell lysate 10 μg (Input).
Lane 2 : ab245116 IP in THP-1 whole cell lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab245116 in THP-1 whole cell lysate (-).

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 minute.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245116).

All lanes:

Immunoprecipitation - Anti-Flt3 / CD135 antibody [EPR22232-318] (<a href='/en-us/products/primary-antibodies/flt3-cd135-antibody-epr22232-318-ab245116'>ab245116</a>)

Predicted band size: 112 kDa

Observed band size: 120 kDa

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Supplier Data

Western blot - Anti-Flt3 / CD135 antibody [EPR22232-318] - BSA and Azide free (AB245824)

False colour image of Western blot : Anti-Flt3 / CD135 antibody [EPR22232-318] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab245116 was shown to bind specifically to Flt3 / CD135. A band was observed at 130 kDa in wild-type THP-1 cell lysates with no signal observed at this size in FLT3 knockout cell line ab282857 (knockout cell lysate ab283225). The band observed at 60 kDa has not been identified. To generate this image, wild-type and FLT3 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245116).

All lanes:

Western blot - Anti-Flt3 / CD135 antibody [EPR22232-318] (<a href='/en-us/products/primary-antibodies/flt3-cd135-antibody-epr22232-318-ab245116'>ab245116</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human FLT3 (CD135) knockout THP-1 cell lysate (ab283225) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 112 kDa

Observed band size: 130 kDa

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Unconjugated

Anti-Flt3 / CD135 antibody [EPR22232-318]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR22232-318

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB, IP

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"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }

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Product details

ab245824 is the carrier-free version of ab245116.

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

Storage information

Do Not Freeze

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Supplementary information

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

Flt3 also known as CD135 or FLT3 tyrosine kinase is a receptor tyrosine kinase that belongs to the class III receptor tyrosine kinase family. It has a molecular weight of approximately 160 kDa. This protein plays an important role in the development of hematopoietic stem and progenitor cells. Flt3 is expressed mainly in early progenitor cells within the bone marrow. Additionally it is present in some populations of mature cells in the peripheral blood and lymphoid organs.

Biological function summary

Flt3 functions as a receptor that is activated by binding to its ligand FLT3 ligand (FLT3L). This interaction stimulates the associated tyrosine kinase activity leading to auto-phosphorylation of Flt3 and subsequent downstream signaling. Flt3 does not form a heterocomplex rather it dimerizes upon ligand binding to initiate signaling cascades. These cascades promote cell proliferation differentiation and survival particularly impacting hematopoiesis and immune responses.

Pathways

Flt3 signaling impacts the internal signaling networks involving the MAPK/ERK and PI3K/AKT pathways both central to cell cycle regulation and apoptosis. Flt3 interacts functionally with proteins such as SHP2 and GRB2 in these pathways aiding signal transduction. The Flt3 receptor also shares functional motifs with KIT and PDGF receptors indicating shared regulatory mechanisms involved in cellular proliferation.

Flt3 mutations especially internal tandem duplications in Flt3/ITD are significant in acute myeloid leukemia (AML) leading to abnormal cell growth and survival. These mutations often co-occur with other genetic anomalies such as NPM1 mutations contributing to the oncogenic process. The abnormal activation of Flt3 kinase activity also has ramifications in myelodysplastic syndromes making it a target for therapeutic intervention with drugs such as anti-FLT3 inhibitors in the treatment regime.

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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

Tyrosine-protein kinase that acts as a cell-surface receptor for the cytokine FLT3LG and regulates differentiation, proliferation and survival of hematopoietic progenitor cells and of dendritic cells. Promotes phosphorylation of SHC1 and AKT1, and activation of the downstream effector MTOR. Promotes activation of RAS signaling and phosphorylation of downstream kinases, including MAPK1/ERK2 and/or MAPK3/ERK1. Promotes phosphorylation of FES, FER, PTPN6/SHP, PTPN11/SHP-2, PLCG1, and STAT5A and/or STAT5B. Activation of wild-type FLT3 causes only marginal activation of STAT5A or STAT5B. Mutations that cause constitutive kinase activity promote cell proliferation and resistance to apoptosis via the activation of multiple signaling pathways.

See full target information FLT3

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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-Flt3 / CD135 antibody [EPR22232-318] - BSA and Azide free

Immunoprecipitation - Anti-Flt3 / CD135 antibody [EPR22232-318] - BSA and Azide free (AB245824)

Western blot - Anti-Flt3 / CD135 antibody [EPR22232-318] - BSA and Azide free (AB245824)

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Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

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Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Product promise