Name: Anti-IRF5 antibody [10T1]
SKU: ab33478
Rating: 4 (3 reviews)

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Western blot - Anti-IRF5 antibody [10T1] (ab33478)

Key features and details

Mouse monoclonal [10T1] to IRF5
Suitable for: Flow Cyt (Intra), ICC/IF, WB
Knockout validated
Reacts with: Mouse, Human
Isotype: IgG1

製品名

Anti-IRF5 antibody [10T1]
IRF5 一次抗体製品一覧
製品の詳細

Mouse monoclonal [10T1] to IRF5
由来種

Mouse
アプリケーション

適用あり: Flow Cyt (Intra), ICC/IF, WBmore details
種交差性

交差種: Mouse, Human
免疫原

Recombinant human IRF-5 (176-240 aa, isoform D) purified from E. coli. NCBI Accession No.: NP_001092099 (formally NP_002191).
ポジティブ･コントロール
- WB: Extracts of Ramos, THP-1 and A20 cells. Flow Cyt (Intra): THP-1 cells. ICC/IF: THP-1, Raw264.7 and HEK293.
特記事項

This product was changed from ascites to tissue culture supernatant on 24/01/2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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.
バッファー

pH: 7.40
Preservative: 0.1% Sodium azide
Constituent: 99% PBS
Concentration information loading...
精製度

Protein G purified
ポリ/モノ

モノクローナル
クローン名

10T1
ミエローマ

Sp2/0
アイソタイプ

IgG1
軽鎖の種類

kappa
研究分野
- Neuroscience
- Processes

Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
- Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
Positive Controls
Recombinant Protein
- Recombinant Human IRF5 protein (ab114295)

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ab33478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 (Intra)		Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF		Use at an assay dependent concentration.
WB	(1)	Use at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).

特記事項
Flow Cyt (Intra) Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).

関連疾患

Genetic variations in IRF5 are associated with susceptibility to inflammatory bowel disease type 14 (IBD14) [MIM:612245]. IBD14 is a chronic, relapsing inflammation of the gastrointestinal tract with a complex etiology. It is subdivided into Crohn disease and ulcerative colitis phenotypes. Crohn disease may affect any part of the gastrointestinal tract from the mouth to the anus, but most frequently it involves the terminal ileum and colon. Bowel inflammation is transmural and discontinuous; it may contain granulomas or be associated with intestinal or perianal fistulas. In contrast, in ulcerative colitis, the inflammation is continuous and limited to rectal and colonic mucosal layers; fistulas and granulomas are not observed. Both diseases include extraintestinal inflammation of the skin, eyes, or joints.
Genetic variations in IRF5 are associated with susceptibility to systemic lupus erythematosus type 10 (SLEB10) [MIM:612251]. Systemic lupus erythematosus (SLE) is a chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. It is thought to represent a failure of the regulatory mechanisms of the autoimmune system.
Genetic variations in IRF5 are a cause of susceptibility to rheumatoid arthritis (RA) [MIM:180300]. It is a systemic inflammatory disease with autoimmune features and a complex genetic component. It primarily affects the joints and is characterized by inflammatory changes in the synovial membranes and articular structures, widespread fibrinoid degeneration of the collagen fibers in mesenchymal tissues, and by atrophy and rarefaction of bony structures.
配列類似性

Belongs to the IRF family.
Contains 1 IRF tryptophan pentad repeat DNA-binding domain.
細胞内局在

Nucleus.
Target information above from: UniProt accession Q13568 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: 3663 Human
- Entrez Gene: 27056 Mouse
- Omim: 607218 Human
- SwissProt: Q13568 Human
- SwissProt: P56477 Mouse
- Unigene: 521181 Human
- Unigene: 6479 Mouse
別名
- Interferon regulatory factor 5 antibody
- Interferon regulatory factor 5 bone marrow variant antibody
- IRF 5 antibody
- IRF-5 antibody
- Irf5 antibody
- IRF5_HUMAN antibody
- SLEB10 antibody
see all

Western blot - Anti-IRF5 antibody [10T1] (ab33478)

All lanes : Anti-IRF5 antibody [10T1] (ab33478) at 1/500 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : IRF5 knockout HAP1 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 58 kDa

Lanes 1 - 2: Merged signal (red and green). Green - ab33478 observed at 56 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab33478 was shown to specifically react with IRF5 in wild-type HAP1 cells as signal was lost in IRF5 knockout cells. Wild-type and IRF5 knockout samples were subjected to SDS-PAGE. Ab33478 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

This image was produced using the ascites version of this antibody.
Flow Cytometry (Intracellular) - Anti-IRF5 antibody [10T1] (ab33478)

Flow cytometry analysis of IRF5 in THP-1 cell line, staining at 2-5µg for 1x10⁶cells (red line). The secondary antibody used goat anti-mouse IgG Alexa fluor 488 conjugate. Isotype control antibody was mouse IgG (black line).
Immunocytochemistry/ Immunofluorescence - Anti-IRF5 antibody [10T1] (ab33478)

Immunocytochemistry/ Immunofluorescence analysis of IRF5 in Raw264.7 cells. The cell was stained with ab33478 (1:100). The secondary antibody (green) was used Alexa Fluor 488. DAPI was stained the cell nucleus (blue).
Immunocytochemistry/ Immunofluorescence - Anti-IRF5 antibody [10T1] (ab33478)

Immunocytochemistry/ Immunofluorescence analysis of IRF5 in THP-1 cells. The cell was stained with ab33478 (1:100). The secondary antibody (green) was used Alexa Fluor 488. DAPI was stained the cell nucleus (blue).
Western blot - Anti-IRF5 antibody [10T1] (ab33478)

Anti-IRF5 antibody [10T1] (ab33478) at 1/1000 dilution + THP-1 cell lysate

Secondary
Goat anti-mouse secondary antibody conjugated to HRP

Predicted band size: 58 kDa
Immunocytochemistry/ Immunofluorescence - Anti-IRF5 antibody [10T1] (ab33478)

Immunofluorescence/Immunocytochemistry analysis of HEK293 cells (10⁵-10⁶) stained with Mouse monoclonal [10T1] to IRF5 (ab33478 1/100-1/200).

This image was produced using the ascites version of this antibody.
Western blot - Anti-IRF5 antibody [10T1] (ab33478)

All lanes : Anti-IRF5 antibody [10T1] (ab33478) at 1/1000 dilution

Lane 1 : Ramos cell lysate
Lane 2 : THP-1 cell lysate
Lane 3 : A20 cell lysate
Lane 4 : NIH 3T3 cell lysate

Secondary
All lanes : goat anti-mouse secondary antibody conjugated to HRP

Developed using the ECL technique.

Predicted band size: 58 kDa
Observed band size: 58 kDa
Additional bands at: 26 kDa, 80 kDa. We are unsure as to the identity of these extra bands.

100-200µg of whole cell lysates were used for western blotting.

This image was produced using the ascites version of this antibody.
Flow Cytometry (Intracellular) - Anti-IRF5 antibody [10T1] (ab33478)

Overlay histogram showing THP1 cells stained with ab33478 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab33478, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in THP1 cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

This image was produced using the ascites version of this antibody.

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

ab33478 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。

ab33478 は 16 報の論文で使用されています。

Moen SH et al. Human Toll-like Receptor 8 (TLR8) Is an Important Sensor of Pyogenic Bacteria, and Is Attenuated by Cell Surface TLR Signaling. Front Immunol 10:1209 (2019). PubMed: 31214180
Guo J et al. A promising role of interferon regulatory factor 5 as an early warning biomarker for the development of human non-small cell lung cancer. Lung Cancer 135:47-55 (2019). PubMed: 31447002
Xiao Y et al. An oligodeoxynucleotide with AAAG repeats significantly attenuates burn-induced systemic inflammatory responses via inhibiting interferon regulatory factor 5 pathway. Mol Med 23:N/A (2017). PubMed: 28620671
Koelzer VH et al. Digital analysis and epigenetic regulation of the signature of rejection in colorectal cancer. Oncoimmunology 6:e1288330 (2017). PubMed: 28507795
Chen X et al. MicroRNA-302a suppresses influenza A virus-stimulated interferon regulatory factor-5 expression and cytokine storm induction. J Biol Chem 292:21291-21303 (2017). PubMed: 29046356

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

1-5 of 5 Abreviews or Q&A

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Macrophages)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Ventana Ultra CC1

Permeabilization

Specification

Macrophages

Blocking step

OptiView H2O2 as blocking agent for 8 minute(s) · Concentration: 0.04% · Temperature: 36°C

Fixative

Formaldehyde

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.

Abcam user community

Verified customer

投稿 Jan 25 2016

Application

Western blot

Loading amount

30 µg

Gel Running Conditions

Reduced Denaturing

Sample

Human Cell lysate - whole cell (macrophage)

Specification

macrophage

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

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.

Abcam user community

Verified customer

投稿 Dec 12 2014

Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (Brain)

Specification

Brain

Fixative

Paraformaldehyde

Permeabilization

Yes - heat, Triton X-100

Blocking step

Sniper (BioCare) as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 25°C

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.

MS. Colleen Karlo

Verified customer

投稿 Oct 04 2011

Question

Phone call reporting problems in using anti-IRF5 antibody [10T1] (ab33478) in western blotting.

Abcam community

Verified customer

Asked on Apr 18 2012

Answer

Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As we discussed on the phone, I have attachedour questionnaire so that we can gather further information regarding the samples tested and the protocol used.This information will also allow us to investigate this case internally and initiate additional testing where necessary.

As discussed over the phone, I have arranged for a replacement vial of ab33478 to be sent out to you. This should reach you tomorrow or Friday. The order number is xxxxxx (Purchase order number FOCR xxxxx). If you do not find that your results improve with this new lot please do let me know.

I look forward to receiving your reply.

Abcam Scientific Support

Answered on Apr 18 2012

Question

I have a few questions about your anti-IRF5 antibody, cat.#:ab33478. Q1: what is the suggested concentration for detection of IRF5 in human dendritic cells? Q2: In the web site, the band shown in lane 2 is the endogenous IRF5 or inducible IRF5 in THP-1 cells? If it is inducible, what is the inducer and the concentration? Q3: Is ab6728 is the cat.# for HRP antibody used to show the IRF5 band? Q4: Dose milk or BSA used in blocking buffer make any differences in showing IFR5 expression? I would greatly appreciate your promptly response.

Abcam community

Verified customer

Asked on Jul 17 2006

Answer

Thank you for your interest in our product, anti-IRF5 antibody(ab33478). I now have some more details which should answer your questions. 1. The anti-IRF5 Antibody (ab33478) was used at 1:1,000 dilution, and 30~50mg protein of cell lysates was used for detection in western blotting. 2. Bands shown in the data sheet are endogenous IRF5, they are not inducible protein. 3. A commercialised secondary antibody, goat anti-mouse Ig-HRP conjugated was used. This was not ab6728. 4. You can get same results for milk or BSA as blocking solution, So you may use anything. I hope that theses answers are helpful for you and please contact me again if you have any other questions. With thanks.

Abcam Scientific Support

Answered on Jul 19 2006

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

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

バッファー

精製度

ポリ/モノ

クローン名

ミエローマ

アイソタイプ

軽鎖の種類

研究分野

関連製品

Compatible Secondaries

Conjugation kits

Isotype control

Positive Controls

Recombinant Protein

アプリケーション

The Abpromise guarantee

ターゲット情報

関連疾患

配列類似性

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 (16)

レビューと Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-IRF5 antibody [10T1]

Western blot abreview for Anti-IRF5 antibody [10T1]

Immunohistochemistry (Frozen sections) abreview for Anti-IRF5 antibody [10T1]