Name: Anti-ICAM1 antibody [EPR24639-3]
Brand: RabMAb
SKU: ab282575
Rating: 5 (1 reviews)

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Western blot - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR24639-3] to ICAM1
Suitable for: Flow Cyt, IP, IHC-P, ICC/IF, WB
Knockout validated
Reacts with: Rat, Human

Carrier Free

製品名

Anti-ICAM1 antibody [EPR24639-3]
ICAM1 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR24639-3] to ICAM1
由来種

Rabbit
特異性

Rat species is recommended based on Flow Cyt result. We do not guarantee other applications for rat.
アプリケーション

適用あり: Flow Cyt, IP, IHC-P, ICC/IF, WBmore details
適用なし: IHC-Fr
種交差性

交差種: Rat, Human
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- WB: Wild-type HeLa whole cell lysate. Ramos and Raji whole cell lysate. Human kidney tissue lysate. IHC-P: Human kidney tissue. Human breast cancer tissue. ICC/IF: Raji cells. Wild-type HeLa cells. Flow Cyt: Rat splenocytes. C6 and Raji cells. IP: Raji whole cell 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. Avoid freeze / thaw cycle.
バッファー

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59.94% 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

EPR24639-3
アイソタイプ

IgG
研究分野
- Neuroscience
- Development

Alternative Versions
- Anti-ICAM1 antibody [EPR24639-3] - BSA and Azide free (ab282599)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human ICAM1 knockout HeLa cell line (ab261742)
KO cell lysates
- Human ICAM1 knockout HeLa cell lysate (ab256947)

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ab282575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
Flow Cyt		1/500.
IP		1/30.
IHC-P		1/600. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/500.
WB	(1)	1/1000. Predicted molecular weight: 57 kDa.

特記事項
Flow Cyt 1/500.
IP 1/30.
IHC-P 1/600. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
WB 1/1000. Predicted molecular weight: 57 kDa.

追加情報

Is unsuitable for IHC-Fr.

機能

ICAM proteins are ligands for the leukocyte adhesion protein LFA-1 (integrin alpha-L/beta-2). During leukocyte trans-endothelial migration, ICAM1 engagement promotes the assembly of endothelial apical cups through ARHGEF26/SGEF and RHOG activation. In case of rhinovirus infection acts as a cellular receptor for the virus.
配列類似性

Belongs to the immunoglobulin superfamily. ICAM family.
Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
翻訳後修飾

Monoubiquitinated, which is promoted by MARCH9 and leads to endocytosis.
細胞内局在

Membrane.
Target information above from: UniProt accession P05362 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: 3383 Human
- Entrez Gene: 25464 Rat
- Omim: 147840 Human
- SwissProt: P05362 Human
- SwissProt: Q00238 Rat
- Unigene: 643447 Human
- Unigene: 12 Rat
別名
- Antigen identified by monoclonal antibody BB2 antibody
- BB 2 antibody
- BB2 antibody
- CD 54 antibody
- CD_antigen=CD54 antibody
- CD54 antibody
- Cell surface glycoprotein P3.58 antibody
- Human rhinovirus receptor antibody
- ICAM 1 antibody
- ICAM-1 antibody
- ICAM1 antibody
- ICAM1_HUMAN antibody
- intercellular adhesion molecule 1 (CD54) antibody
- intercellular adhesion molecule 1 (CD54), human rhinovirus receptor antibody
- Intercellular adhesion molecule 1 antibody
- Major group rhinovirus receptor antibody
- MALA 2 antibody
- MALA2 antibody
- MyD 10 antibody
- MyD10 antibody
- P3.58 antibody
- Surface antigen of activated B cells antibody
- Surface antigen of activated B cells, BB2 antibody
see all

Western blot - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

All lanes : Anti-ICAM1 antibody [EPR24639-3] (ab282575) at 1/1000 dilution

Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ICAM1 knockout HeLa whole cell lysate
Lane 3 : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution

Predicted band size: 57 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS

Lanes 1-3: Merged signal (red and green). Green - ab282575 observed at 90kDa. Red - loading control ab8245 observed at 36 kDa.

ab282575 Anti-ICAM1 antibody [EPR24639-3] was shown to react with ICAM1 in wild-type Hela cells in Western blot. Loss of signal was observed when knockout cell line ab261742 (knockout cell lysate ab256947) was used. Wild-type and ICAM1 knockout samples were subjected to SDS-PAGE.

ab282575 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight 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 10000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

All lanes : Anti-ICAM1 antibody [EPR24639-3] (ab282575) at 1/1000 dilution

Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ICAM1 knockout Hela whole cell lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 57 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?

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

ab282575 Anti-ICAM1 antibody [EPR24639-3] was shown to react with ICAM1 in wild-type Hela cells in Western blot. Loss of signal was observed when knockout cell line ab261742 (knockout cell lysate ab256947) was used. Wild-type and ICAM1 knockout samples were subjected to SDS-PAGE.

Exposure time: 15 seconds.
Western blot - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

Anti-ICAM1 antibody [EPR24639-3] (ab282575) at 1/1000 dilution + human kidney tissue lysate at 20 µg

Secondary
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

Predicted band size: 57 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?

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

Exposure time: 26 seconds
Western blot - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

All lanes : Anti-ICAM1 antibody [EPR24639-3] (ab282575) at 1/1000 dilution

Lane 1 : Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 2 : Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 57 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?

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

Exposure time: 15 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labelling ICAM1 with ab282575 at 1/600 (0.902 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on normal human glomerulus. The section was incubated with ab282575 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labelling ICAM1 with ab282575 at 1/600 (0.902 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human breast cancer (PMID: 30082828). The section was incubated with ab282575 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji cells labelling ICAM1 with ab282575 at 1/500 (1.082 µg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in Raji cells.

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized ICAM1 KO HeLa cells labelling ICAM1 with ab282575 at 1/500 (1.082 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in wild-type HeLa cells, and no staining in ICAM1 knockout HeLa cells is observed.

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.
Flow Cytometry - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

Flow cytometric analysis of rat splenocytes cells labelling ICAM1 with ab282575 at 1/500 dilution (0.1 µg)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.
Flow Cytometry - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

Flow cytometric analysis of C6 (Rat glial tumor glial cell) cells labelling ICAM1 with ab282575 at 1/500 dilution (0.1 µg)/(red) compared with a Rabbit monoclonal IgG (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, ab150077) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.
Flow Cytometry - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

Flow cytometric analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labelling ICAM1 with ab282575 at 1/500 dilution (0.1 µg)/(Red) compared with a Rabbit monoclonal IgG (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, ab150077) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.
Immunoprecipitation - Anti-ICAM1 antibody [EPR24639-3] (ab282575)

ICAM1 was immunoprecipitated from 0.35 mg Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate 10 µg with ab282575 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab282575 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Raji whole cell lysate 10 µg

Lane 2: ab282575 IP in Raji whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab282575 in Raji whole cell lysate

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

Exposure time: 3 minutes.
Anti-ICAM1 antibody [EPR24639-3] (ab282575)

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

Language
Datasheet download

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ab282575 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。

ab282575 は 10 報の論文で使用されています。

Zhang J et al. Shensong Yangxin Capsule Reduces the Susceptibility of Arrhythmia in db/db Mice via Inhibiting the Inflammatory Response Induced by Endothelium Dysfunction. Drug Des Devel Ther 17:313-330 (2023). PubMed: 36776448
Li J et al. A novel inflammation-related signature for predicting prognosis and characterizing the tumor microenvironment in colorectal cancer. Aging (Albany NY) 15:2554-2581 (2023). PubMed: 37014331
Zhang M et al. Exercise Training Attenuates Acute β-Adrenergic Receptor Activation-Induced Cardiac Inflammation via the Activation of AMP-Activated Protein Kinase. Int J Mol Sci 24:N/A (2023). PubMed: 37298222
Wang Y et al. Differentially expressed genes in orbital adipose/connective tissue of thyroid-associated orbitopathy. PeerJ 11:e16569 (2023). PubMed: 38130930
Chen Y et al. Taxifolin improves inflammatory injury of human bronchial epithelial cells by inhibiting matrix metalloproteinase (MMP) 10 via Wnt/β-catenin pathway. Bioengineered 13:1198-1208 (2022). PubMed: 35000533
Tan Z et al. KLF2 alleviates endothelial cell injury and inhibits the formation of THP‑1 macrophage‑derived foam cells by activating Nrf2 and enhancing autophagy. Exp Ther Med 24:737 (2022). PubMed: 36478888
Lin J et al. Transcript Engineered Extracellular Vesicles Alleviate Alloreactive Dynamics in Renal Transplantation. Adv Sci (Weinh) 9:e2202633 (2022). PubMed: 36073846
Song X et al. Semaphorin 7A knockdown improves injury and prevents endothelial-to-mesenchymal transition in ox-LDL-induced HUVECs by regulating β1 integrin expression. Exp Ther Med 22:1441 (2021). PubMed: 34721683
Huang HW et al. Fibroblast Growth Factor Type 1 Ameliorates High-Glucose-Induced Oxidative Stress and Neuroinflammation in Retinal Pigment Epithelial Cells and a Streptozotocin-Induced Diabetic Rat Model. Int J Mol Sci 22:N/A (2021). PubMed: 34281287
Yan Z et al. Cancer-associated fibroblast-derived exosomal miR-18b promotes breast cancer invasion and metastasis by regulating TCEAL7. Cell Death Dis 12:1120 (2021). PubMed: 34853307

レビューと Q&A

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Application

Western blot

Sample

Cynomolgus monkey Tissue lysate - whole (Lung)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

5 µg

Specification

Lung

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 10%

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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投稿 Aug 25 2023

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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

特異性

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

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

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

配列類似性

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

Certificate of Compliance

参考文献 (10)

レビューと Q&A

Western blot abreview for Anti-ICAM1 antibody [EPR24639-3]