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Rabbit Recombinant Monoclonal CARD9 antibody. Suitable for Flow Cyt (Intra), WB and reacts with Human, Mouse samples.

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Images

Flow Cytometry (Intracellular) - Anti-CARD9 antibody [EPR28960-91] (AB322352), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-CARD9 antibody [EPR28960-91] (AB322352), expandable thumbnail
  • Western blot - Anti-CARD9 antibody [EPR28960-91] (AB322352), expandable thumbnail
  • Western blot - Anti-CARD9 antibody [EPR28960-91] (AB322352), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow Cyt (Intra)WBICC/IFIHC-P
Human
Tested
Tested
Not recommended
Not recommended
Mouse
Tested
Tested
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
1/5000
Notes

-

Species
Mouse
Dilution info
1/5000
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Species
Mouse
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Human, Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse
Dilution info
-
Notes

-

Target data

Function

Adapter protein that plays a key role in innate immune response against fungi by forming signaling complexes downstream of C-type lectin receptors (PubMed:26961233, PubMed:33558980). CARD9-mediated signals are essential for antifungal immunity against a subset of fungi from the phylum Ascomycota (PubMed:24231284, PubMed:25057046, PubMed:25702837, PubMed:26521038, PubMed:26679537, PubMed:26961233, PubMed:27777981, PubMed:29080677, PubMed:33558980). Transduces signals in myeloid cells downstream of C-type lectin receptors CLEC7A (dectin-1), CLEC6A (dectin-2) and CLEC4E (Mincle), which detect pathogen-associated molecular pattern metabolites (PAMPs), such as fungal carbohydrates, and trigger CARD9 activation (By similarity). Upon activation, CARD9 homooligomerizes to form a nucleating helical template that recruits BCL10 via CARD-CARD interaction, thereby promoting polymerization of BCL10 and subsequent recruitment of MALT1: this leads to activation of NF-kappa-B and MAP kinase p38 (MAPK11, MAPK12, MAPK13 and/or MAPK14) pathways which stimulate expression of genes encoding pro-inflammatory cytokines and chemokines (PubMed:11053425, PubMed:26488816, PubMed:26961233, PubMed:31296852, PubMed:33558980). CARD9 signaling in antigen-presenting cells links innate sensing of fungi to the activation of adaptive immunity and provides a cytokine milieu that induces the development and subsequent of interleukin 17-producing T helper (Th17) cells (PubMed:24231284). Also involved in activation of myeloid cells via classical ITAM-associated receptors and TLR: required for TLR-mediated activation of MAPK, while it is not required for TLR-induced activation of NF-kappa-B (By similarity). CARD9 can also be engaged independently of BCL10: forms a complex with RASGRF1 downstream of C-type lectin receptors, which recruits and activates HRAS, leading to ERK activation and the production of cytokines (By similarity). Acts as an important regulator of the intestinal commensal fungi (mycobiota) component of the gut microbiota (PubMed:33548172). Plays an essential role in antifungal immunity against dissemination of gut fungi: acts by promoting induction of antifungal IgG antibodies response in CX3CR1(+) macrophages to confer protection against disseminated C.albicans or C.auris infection (PubMed:33548172). Also mediates immunity against other pathogens, such as certain bacteria, viruses and parasites; CARD9 signaling is however redundant with other innate immune responses (By similarity). In response to L.monocytogenes infection, required for the production of inflammatory cytokines activated by intracellular peptidoglycan: acts by connecting NOD2 recognition of peptidoglycan to downstream activation of MAP kinases (MAPK) without activating NF-kappa-B (By similarity).

Alternative names

Caspase recruitment domain-containing protein 9, hCARD9, CARD9

Rabbit Recombinant Monoclonal CARD9 antibody. Suitable for Flow Cyt (Intra), WB and reacts with Human, Mouse samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR28960-91
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

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

Notes

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

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Flow Cytometry (Intracellular) - Anti-CARD9 antibody [EPR28960-91] (ab322352), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CARD9 antibody [EPR28960-91] (ab322352)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MEF (mouse embryo fibroblast, Left)/ RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Right) cells labelling CARD9 with ab322352 at 1/5000 (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at 1/5000 was used as the secondary antibody.

    Negative Control: MEF.

  • Flow Cytometry (Intracellular) - Anti-CARD9 antibody [EPR28960-91] (ab322352), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CARD9 antibody [EPR28960-91] (ab322352)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A549 (human lung carcinoma epithelial cell, Left) / THP-1 (human monocytic leukemia monocyte, Right) cells labelling CARD9 with ab322352 at 1/5000 (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at 1/5000 was used as the secondary antibody.

    Low expression: A549.

  • Western blot - Anti-CARD9 antibody [EPR28960-91] (ab322352), expandable thumbnail

    Western blot - Anti-CARD9 antibody [EPR28960-91] (ab322352)

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

    Negative control: MEF.

    Low expression: skeletal muscle (PMID: 11053425).

    The expression profile/ molecular weight observed is consistent with what has been described in the literature(PMID: 32164050), This antibody can recognize 3 isoforms of CARD9 and isoforms have different molecular weight.

    The identity of the lower MW band at about 30 kDa (in lane 1) is unknown.

    The identity of the higher MW bands at about 100 kDa and the lower MW bands at about 40 kDa (in lane 4) are unknown.

    This blot of lanes 3-4 was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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

    All lanes: Western blot - Anti-CARD9 antibody [EPR28960-91] (ab322352) at 1/1000 dilution

    Lane 1: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 40 µg

    Lane 2: MEF (mouse embryo fibroblast) whole cell lysate at 40 µg

    Lane 3: Mouse spleen tissue lysate at 40 µg

    Lane 4: Mouse skeletal muscle lysate at 40 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 43-62 kDa, 36 kDa

    Exposure time: 136s

  • Western blot - Anti-CARD9 antibody [EPR28960-91] (ab322352), expandable thumbnail

    Western blot - Anti-CARD9 antibody [EPR28960-91] (ab322352)

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

    Low expression: A549 (PMID: 32164050 ), 293T (PMID: 33558980).

    The expression profile/ molecular weight observed is consistent with what has been described in the literature(PMID: 32164050), This antibody can recognize 3 isoforms of CARD9 and isoforms have different molecular weight.

    The identity of the higher MW band at 150 kDa (in lane 1) is unknown.

    The identity of the lower MW band at 25 kDa (in lane 5) is unknown.

    This blot of lane 1 and lane 5 was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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

    All lanes: Western blot - Anti-CARD9 antibody [EPR28960-91] (ab322352) at 1/1000 dilution

    Lane 1: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

    Lane 2: THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

    Lane 3: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Lane 5: Human bone marrow tissue lysate at 40 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 43-62 kDa, 36 kDa

    Exposure time: 180s

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Product protocols

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

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