製品の概要

  • 製品名
    Anti-Involucrin antibody [SY5]
    Involucrin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [SY5] to Involucrin
  • 由来種
    Mouse
  • アプリケーション
    適用あり: ICC/IF, WB, IP, IHC-P, IHC-Fr, Flow Cytmore details
  • 種交差性
    交差種: Dog, Human, Pig, Gorilla, Owl monkey
    非交差種: Mouse
  • 免疫原

    Pure involucrin from human keratinocytes.

  • エピトープ
    The epitope maps between codon 421-568 of human involucrin.
  • ポジティブ・コントロール
    • IHC-P: Human tonsil, skin tissue. Localized to upper spinous and granular layers in normal skin. Flow cyt: A431 cells. ICC/IF: MCF7 cells.
  • 特記事項

    Involucrin is the differentiation marker of human keratinocytes.
    ab81216 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

製品の特性

  • 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 or -80°C. Avoid freeze / thaw cycle.
  • バッファー
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 精製度
    Immunogen affinity purified
  • ポリ/モノ
    モノクローナル
  • クローン名
    SY5
  • ミエローマ
    P3x63
  • アイソタイプ
    IgG1
  • 軽鎖の種類
    unknown
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab68 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 1 µg/ml.
WB Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 0.1 - 0.2 µg/ml.
IHC-Fr 1/100.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能
    Part of the insoluble cornified cell envelope (CE) of stratified squamous epithelia.
  • 組織特異性
    Keratinocytes of epidermis and other stratified squamous epithelia.
  • 配列類似性
    Belongs to the involucrin family.
  • 翻訳後修飾
    Substrate of transglutaminase. Some glutamines and lysines are cross-linked to other involucrin molecules, to other proteins such as keratin, desmoplakin, periplakin and envoplakin, and to lipids like omega-hydroxyceramide.
  • 細胞内局在
    Cytoplasm. Constituent of the scaffolding of the cornified envelope.
  • Information by UniProt
  • 参照データベース
  • 別名
    • INVO_HUMAN antibody
    • Involucrin antibody
    • IVL antibody

画像

  • ab68 at a 1/1000 dilution staining Involucrin in human normal skin tissue by Immunohistochemistry (formalin fixed, paraffin embedded sections), incubated for 2 hours at 21°C. Heat mediated antigen retrieval step performed using citrate buffer pH 6.0. Blocked with 10% serum for 30 minutes at 21°C. Secondary used at 1/100 polyclonal Donkey anti-mouse IgG conjugated to Alexa Fluor 594 (red).

    See Abreview

  • KLC (keratinocyte-like cells) have the capacity to stratify

    Human skin was decellularized and confirmed complete cell removal by H&E staining. ASC (human adipose-derived stem cells) were seeded on the epithelial side of human dermal decellularized matrices in the presence or absence of EGF (0.005 µg/mL). Human dermal decellularized matrices seeded with keratinocytes were used as controls. All three groups were kept in the same media conditions and lifted to an air-liquid interphase at the same time. Histological analysis revealed the presence of several layers of KLC with an organization similar to that of keratinocytes (A.1and 3). Surprisingly, ASC without EGF not only did not transdifferentiate into KLC, but instead migrated inside the matrix (A.2). Immunohistochemical analysis revealed a positive staining of cytokeratin-5 (B.3), cytokeratin-10 (C.3) and Involucrin (D.3) in stratified KLC matrices comparable to that of the stratified keratinocyte matrix (A, B, C and D.1). No positive staining was found in the ASC seeded matrices (A, B, C and D.2).

  • Overlay histogram showing A431 cells stained with ab68 (red line). The cells were fixed with 80% methanol (5 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 followed by the antibody (ab68, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H&L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • ICC/IF image of ab68 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab68, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunohistochemical analysis of Formalin fixed paraffin embedded human tonsil section labelling Involucrin with ab68 at dilution of 0.1 ug/mL. 

  • Immunohistochemical analysis of frozen Human artifical skin tissue labeling Involucrin with ab68 at 1/200 dilution.

    See Abreview

参考文献

This product has been referenced in:
  • Dziunycz PJ  et al. CYFIP1 is directly controlled by NOTCH1 and down-regulated in cutaneous squamous cell carcinoma. PLoS One 12:e0173000 (2017). WB ; Human . Read more (PubMed: 28410392) »
  • Davies EG  et al. Thymus transplantation for complete DiGeorge syndrome: European experience. J Allergy Clin Immunol 140:1660-1670.e16 (2017). IHC-P ; Human . Read more (PubMed: 28400115) »
See all 24 Publications for this product

レビューと Q&A

1-10 of 14 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin)
Specification
Skin
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
Acetone

Abcam user community

Verified customer

投稿 Jul 14 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (artificial skin)
Permeabilization
Yes - Triton X-100
Specification
artificial skin
Blocking step
Blocking One as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

投稿 Aug 20 2015

Answer

Thank you for your patience.
I apologize for the delay, but I only heard back from the supplying lab today with the following information:

"Wehave unfortunately no data to explain the differeing bands seen against human tissue.

The antibody was tested against mouse but showed no reactivity."

I hope this information is nevertheless helpful to you.
If you are not satisfied with the antibody, I'd be happy to replace, credit or refund it - if it was used in a tested species or application.Please do let me know.

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

Read More

Answer

I am very sorry for the delayed response.

Despite numerous reminders the source of the antibody only replied to me today with the following information:

"We have no other information other than the codon sequence on the epitope for this antibody.
We are not able to supply the protein but should that ever be possible we will get in touch."

I am sorry I could not be more helpful on this occasion. Please do not hesitate to contact me again if you have further questions.

Read More

Answer

Thank you for contacting us.

I apologize for the wait, but I have not yet heard back from the lab. I have sent another email to follow up with them.

Thank you for your patience and understanding.

Read More

Answer

Thank you for contacting us.

I have contacted the lab for the questions you had, and will let you know their answer.

The database links for human and mouse proteins are as follows:

human: http://www.uniprot.org/uniprot/P07476
mouse: http://www.uniprot.org/uniprot/P48997

I hope this information is of some help to you.
In the meantime, 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

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Skin)
Specification
Skin
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Mr. Carl Hobbs

Verified customer

投稿 Mar 19 2012

Answer

Thank you for contacting us.

I have attached the MSDS for ab68.

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

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin)
Specification
Skin
Fixative
Acetone
Permeabilization
No
Blocking step
Casein as blocking agent for 10 minute(s) · Concentration: 0.25% · Temperature: 20°C

Dr. Vaughan Feisst

Verified customer

投稿 Oct 23 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Normal Skin)
Specification
Normal Skin
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrat, pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 21°C

Abcam user community

Verified customer

投稿 Feb 19 2010

1-10 of 14 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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