Anti-Caldesmon/CDM 抗体 [E89] (ab32330)

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Western blot - Anti-Caldesmon/CDM antibody [E89] (ab32330)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E89] to Caldesmon/CDM
Suitable for: WB, IHC-P, Flow Cyt (Intra), IP, ICC/IF
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free HRP PE

製品名

Anti-Caldesmon/CDM antibody [E89]
Caldesmon/CDM 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [E89] to Caldesmon/CDM
由来種

Rabbit
アプリケーション

適用あり: WB, IHC-P, Flow Cyt (Intra), IP, ICC/IFmore details
種交差性

交差種: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- WB: HeLa and NIH/3T3 cell lysates; IHC-P: Human breast carcinoma, Human leiomyoma and mouse kidney tissue; ICC/IF: NIH/3T3 cells; IP: NIH/3T3 whole cell lysate; Flow Cyt (Intra): NIH/3T3 cells. WB: HeLa, NIH/3T3 whole cell lysates. Rat Liver lysate.
特記事項
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.

製品の状態

Liquid
保存方法

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.
解離定数（K_D 値）

K_D = 1.25 x 10 ^-10 M
Learn more about K_D
バッファー

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

E89
アイソタイプ

IgG
研究分野

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human CALD1 (Caldesmon/CDM) knockout HeLa cell line (ab265026)
KO cell lysates
- Human CALD1 (Caldesmon/CDM) knockout HeLa cell lysate (ab257375)
Positive Controls
- HeLa whole cell lysate (ab29545)
- NIH/3T3 whole cell lysate (ab7179)
Recombinant Protein
- Recombinant Human Caldesmon/CDM protein (ab114302)
Substrate reagent
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab32330の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
WB		1/10000 - 1/20000. Detects a band of approximately 70 kDa (predicted molecular weight: 93 kDa).
IHC-P		1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)		1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP		1/20.
ICC/IF		1/50. For unpurified use at 1/250 - 1/500.

特記事項
WB 1/10000 - 1/20000. Detects a band of approximately 70 kDa (predicted molecular weight: 93 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra) 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP 1/20.
ICC/IF 1/50. For unpurified use at 1/250 - 1/500.

機能

Actin- and myosin-binding protein implicated in the regulation of actomyosin interactions in smooth muscle and nonmuscle cells (could act as a bridge between myosin and actin filaments). Stimulates actin binding of tropomyosin which increases the stabilization of actin filament structure. In muscle tissues, inhibits the actomyosin ATPase by binding to F-actin. This inhibition is attenuated by calcium-calmodulin and is potentiated by tropomyosin. Interacts with actin, myosin, two molecules of tropomyosin and with calmodulin. Also play an essential role during cellular mitosis and receptor capping.
組織特異性

High-molecular-weight caldesmon (isoform 1) is predominantly expressed in smooth muscles, whereas low-molecular-weight caldesmon (isoforms 2, 3, 4 and 5) are widely distributed in non-muscle tissues and cells. Not expressed in skeletal muscle or heart.
配列類似性

Belongs to the caldesmon family.
ドメイン

The N-terminal part seems to be a myosin/calmodulin-binding domain, and the C-terminal a tropomyosin/actin/calmodulin-binding domain. These two domains are separated by a central helical region in the smooth-muscle form.
翻訳後修飾

In non-muscle cells, phosphorylation by CDK1 during mitosis causes caldesmon to dissociate from microfilaments. Phosphorylation reduces caldesmon binding to actin, myosin, and calmodulin as well as its inhibition of actomyosin ATPase activity. Phosphorylation also occurs in both quiescent and dividing smooth muscle cells with similar effects on the interaction with actin and calmodulin and on microfilaments reorganization.
細胞内局在

Cytoplasm > cytoskeleton. Cytoplasm > myofibril. On thin filaments in smooth muscle and on stress fibers in fibroblasts (nonmuscle).
Target information above from: UniProt accession Q05682 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 800 Human
- Entrez Gene: 109624 Mouse
- Entrez Gene: 25687 Rat
- Omim: 114213 Human
- SwissProt: Q05682 Human
- SwissProt: Q62736 Rat
- Unigene: 490203 Human
別名
- CAD antibody
- CALD 1 antibody
- CALD1 antibody
- CALD1_HUMAN antibody
- Caldesmon 1 antibody
- Caldesmon 1 Isoform 1 antibody
- Caldesmon 1 Isoform 2 antibody
- Caldesmon 1 Isoform 3 antibody
- Caldesmon 1 Isoform 4 antibody
- Caldesmon 1 Isoform 5 antibody
- Caldesmon antibody
- Caldesmon1 antibody
- CDM antibody
- H CAD antibody
- HCAD antibody
- L CAD antibody
- LCAD antibody
- MGC21352 antibody
- NAG22 antibody
see all

Western blot - Anti-Caldesmon/CDM antibody [E89] (ab32330)

All lanes : Anti-Caldesmon/CDM antibody [E89] (ab32330) at 1/10000 dilution (Purified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 3 : Rat liver lysate

Lysates/proteins at 15 µg per lane.

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

Predicted band size: 93 kDa
Western blot - Anti-Caldesmon/CDM antibody [E89] (ab32330)

All lanes : Anti-Caldesmon/CDM antibody [E89] (ab32330) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CALD1 knockout HeLa cell lysate
Lane 3 : NIH/3T3 cell lysate
Lane 4 : HEK-293 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 93 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab32330 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

ab32330 Anti-Caldesmon/CDM antibody [E89] was shown to specifically react with Caldesmon/CDM in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265026 (knockout cell lysate ab257375) was used. Wild-type and Caldesmon/CDM knockout samples were subjected to SDS-PAGE. ab32330 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] (ab32330)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human leiomyoma tissue sections labeling Caldesmon/CDM with purified ab32330 at 1/100 dilution (1.21 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Immunocytochemistry/ Immunofluorescence - Anti-Caldesmon/CDM antibody [E89] (ab32330)

Immunocytochemistry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling Caldesmon/CDM with purified ab32330 at 1/50 dilution (2.4 µg/mL). Cells were fixed in 100% Methanol and permeabilized with 0.1% TritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunoprecipitation - Anti-Caldesmon/CDM antibody [E89] (ab32330)

Purified ab32330 at 1/20 dilution (0.6µg) immunoprecipitating Caldesmon/CDM in NIH/3T3 whole cell lysate.
Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate.
Lane 2 (+): ab32330 + NIH/3T3 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32330 in NIH/3T3 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] (ab32330)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling Caldesmon/CDM with purified ab32330 at 1/100 dilution (1.21 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] (ab32330)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue sections labeling Caldesmon/CDM with purified ab32330 at 1/100 dilution (1.21 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Flow Cytometry (Intracellular) - Anti-Caldesmon/CDM antibody [E89] (ab32330)

Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labelling Caldesmon/CDM with purified ab32330 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Anti-Caldesmon/CDM antibody [E89] (ab32330)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

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Datasheet download

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ab32330 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab32330 は 30 報の論文で使用されています。

Zhang D et al. Local injection of adipose-derived mesenchymal stem cells in silk fibroin solution on the regeneration of lower esophageal sphincter in an animal model of GERD. Front Cell Dev Biol 11:993741 (2023). PubMed: 37077418
Qi F et al. Identification of RECK as a protective prognostic indicator and a tumor suppressor through regulation of the ERK/MAPK signaling pathway in gastric cancer. J Transl Med 21:766 (2023). PubMed: 37904179
Nalluri SM et al. Crosstalk between ERK and MRTF-A signaling regulates TGFβ1-induced epithelial-mesenchymal transition. J Cell Physiol 237:2503-2515 (2022). PubMed: 35224740
Kokate SB et al. Caldesmon controls stress fiber force-balance through dynamic cross-linking of myosin II and actin-tropomyosin filaments. Nat Commun 13:6032 (2022). PubMed: 36229430
Wang Y et al. Plocabulin, a Novel Tubulin Inhibitor, Has Potent Antitumour Activity in Patient-Derived Xenograft Models of Soft Tissue Sarcoma. Int J Mol Sci 23:N/A (2022). PubMed: 35806460

View all Publications for this product

レビューと Q&A

Show All レビュー Q&A

レビューを送る質問を送る

1-6 of 6 Abreviews or Q&A

Question

Thank you for your kind reply.

The customer has also requested some specific instructions or a more precise protocol regarding incubation times and recommended buffers for FACS and IF.

Also, could you please mention a cell type or cell line that might serve as a good negative control for Caldsmon?

Any cells known not to express Cladesmon might do.

Thank you for your kind support.

Regards,

Abcam community

Verified customer

Asked on Feb 20 2013

Answer

I have attached an image of our Flow cytometry results with ab32330.

We used untreated HeLa cells as our negative control and Anisomycin-treated HeLa cells as our positive control. The primary dilution was 1:20. Both were permeabilized according to our standard protocol which is described below:

1. Solutions and Reagents
1.1. 1X PBS

1.2. Blocking buffer: 0.5% BSA in 1X PBS

1.3. 2% paraformaldehyde (1% solution - optional for storing samples)

1.4. 1X FACS permeabilizing solution (BD Biosciences cat. #340973)

1.5. Fluorescently-conjugated secondary antibody (various forms)

2. Protocol
2.1. Collect 1x106 cells/sample.

2.2. Wash cells once with blocking buffer.

2.3. Fix cells with 2% paraformaldehyde and incubate at room temperature for 10 min.

2.4. Wash cells once with blocking buffer.

2.5. Add 0.5 ml 1X FACS permeabilizing solution and incubate at room temperature for 10 min.

2.6. Wash cells once with blocking buffer.

2.7. Incubate cells in blocking buffer for 30 min at room temperature.

2.8. Add primary antibody at the appropriate dilution and incubate for 30 min at room temperature.

2.9. Wash twice with blocking buffer and incubate with fluorescently-conjugated secondary antibody for 30 min at room temperature.

2.10. Wash cells twice with blocking buffer.

2.11. Re-suspend cells in 1X PBS and analyze on flow cytometry. Samples can be kept in 1% paraformaldehyde at 4 °C overnight.

Please let me know if you have any additional questions.

Abcam Scientific Support

Answered on Feb 20 2013

Question

Dear Tech support

Could you please specify what is the formal shelf life of product ab32330?
I know it's guaranteed for 6 month from delivery but how long is good for when stored properly?

Thank you for your kind advise.
Regards,

Abcam community

Verified customer

Asked on Jan 03 2013

Answer

Thank you for your enquiry regarding ab32330.

Generally, antibodies can be stored at -20°C for several years if the storage buffer contains cryoprotective agent and the aliquot size is no smaller than 10 µl. Please always check the product datasheet for storage information.

However, I would like to emphasize that Abcam can only offer a full guarantee called Abpromise within 6 months of purchase.

https://www.abcam.com/index.html?pageconfig=resource&rid=10795

If you need any further assistance in the future, please do not hesitate to contact me.

Abcam Scientific Support

Answered on Jan 03 2013

Question

The antibody is needed for Flow Cytometry in Human or Mouse.
Thank you for your help!

Regards,

Abcam community

Verified customer

Asked on Dec 21 2012

Answer

Thank you for your reply and for kindly providing the further information.

We have one antibody for detection of Caldesmon which has been tested in flow cytometry and mouse, which I think is the one you already found:

Caldesmon antibody [E89] (ab32330)

The sequence we used to raise this antibody has 100% homology with all 6 Caldesmon isoforms. Depending on the level of expression in the customer's sample, they may be able to observe all 6.

Regrettably, this is therefore not specific for the smaller isoforms of the protein in this case and may not be what the customer is looking for.

If you have any further questions, please do not hesitate to contact me.

Abcam Scientific Support

Answered on Dec 21 2012

Question

Is antibody ab32330 caldesmon isoform1 specific? could you please specify what is the isoform specificity and if it's not isoform 1, is there an antibody for this protein?

Abcam community

Verified customer

Asked on Jul 04 2012

Answer

Thank you for contacting us.

Checking the immunogen sequence for this antibody, this antibody is not specific to Caldesmon isoform 1. Based on the sequence, it might crossreact with isoform 1-6 of human Caldesmon protein.

We have two antibodies which react only with the heavy form/ Isoform 1:

ab50016 Anti-Caldesmon antibody [hHCD]

https://www.abcam.com/CaldesmonCDM-antibody-hHCD-ab50016.html (or use the following: https://www.abcam.com/CaldesmonCDM-antibody-hHCD-ab50016.html).

and ab11013 Anti-Caldesmon antibody [CALD 5]

https://www.abcam.com/CaldesmonCDM-antibody-CALD-5-ab11013.html (or use the following: https://www.abcam.com/CaldesmonCDM-antibody-CALD-5-ab11013.html).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Abcam Scientific Support

Answered on Jul 04 2012

Question

I am wondering if this antibody (ab32330) recognize the high-molecular weight caldesmon? As shown on your website there are different isoforms in different tissue and I'd like to get antibody for isoform 1.

Abcam community

Verified customer

Asked on Dec 28 2011

Answer

Thank you for contacting us. Our source for this antibody confirms that the antibody does recognize a 120-150 kDa protein in smooth muscle samples. They do not, have a blot image, however. Please do not hesitate to contact us if you need any more advice or information.

Abcam Scientific Support

Answered on Dec 28 2011

Question

Dear sirs, I have ordered your product no ab 32330, rabbit monoclonal to Caldesmon. In the datasheet it says IgG as an isotype. Is this supposed to be IgG1 or what does it mean? With kind regards

Abcam community

Verified customer

Asked on Dec 19 2011

Answer

Thank you for your patience. The isotype of the anti-Caldesmon antibody is simply rabbit IgG. In immunology, no specific isotype has been determined for rabbits. It is only known for human and mouse. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam Scientific Support

Answered on Dec 19 2011

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

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製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

解離定数（KD 値）

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Recombinant Protein

Substrate reagent

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

組織特異性

配列類似性

ドメイン

翻訳後修飾

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

参考文献 (30)

レビューと Q&A

解離定数（K_D 値）