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Anti-C4d 抗体 (ab87424)

Submit a review Q&A (2)References (1)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C4d antibody (ab87424)
  • Western blot - Anti-C4d antibody (ab87424)
  • Immunocytochemistry/ Immunofluorescence - Anti-C4d antibody (ab87424)

Key features and details

  • Rabbit polyclonal to C4d
  • Suitable for: IHC-P, WB, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-C4 antibody [EPR2990(2)] (ab173577)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-C4d antibody
    C4d 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to C4d

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: IHC-P, WB, ICC/IFmore details

  • 種交差性

    交差種: 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

    • This antibody gave a positive signal in Human Plasma Total Protein Lysate.

  • 特記事項

    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

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 1% BSA, 98.98% PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...

  • 精製度

    Immunogen affinity purified

  • ポリ/モノ

    ポリクローナル

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

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ab87424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
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 193 kDa).
ICC/IF
Use a concentration of 5 µg/ml.
特記事項
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 193 kDa).
ICC/IF
Use a concentration of 5 µg/ml.

ターゲット情報

  • 機能

    C4 plays a central role in the activation of the classical pathway of the complement system. It is processed by activated C1 which removes from the alpha chain the C4a anaphylatoxin. The remaining alpha chain fragment C4b is the major activation product and is an essential subunit of the C3 convertase (C4b2a) and the C5 convertase (C3bC4b2a) enzymes of the classical complement pathway.
    Derived from proteolytic degradation of complement C4, C4a anaphylatoxin is a mediator of local inflammatory process. It induces the contraction of smooth muscle, increases vascular permeability and causes histamine release from mast cells and basophilic leukocytes.

  • 関連疾患

    Defects in C4A are the cause of complement component 4A deficiency (C4AD) [MIM:120810]. A rare defect of the complement classical pathway associated with the development of autoimmune disorders, mainly systemic lupus with or without associated glomerulonephritis.

  • 配列類似性

    Contains 1 anaphylatoxin-like domain.
    Contains 1 NTR domain.

  • 翻訳後修飾

    Prior to secretion, the single-chain precursor is enzymatically cleaved to yield the non-identical chains (alpha, beta and gamma). During activation, the alpha chain is cleaved by C1 into C4a and C4b, and C4b stays linked to the beta and gamma chains. Further degradation of C4b by C1 into the inactive fragments C4c and C4d blocks the generation of C3 convertase.
    N- and O-glycosylated. O-glycosylated with a core 1 or possibly core 8 glycan.

  • 細胞内局在

    Secreted.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • acidic C4 antibody
    • Acidic complement C4 antibody
    • basic C4 antibody
    • basic complement C4 antibody
    • C3 and PZP-like alpha-2-macroglobulin domain-containing protein 2 antibody
    • C3 and PZP-like alpha-2-macroglobulin domain-containing protein 3 antibody
    • C4 antibody
    • C4, chido form antibody
    • C4, Rodgers form antibody
    • C4-1 antibody
    • C4a anaphylatoxin antibody
    • C4A antibody
    • C4A2 antibody
    • C4A3 antibody
    • C4A4 antibody
    • C4A6 antibody
    • C4AD antibody
    • C4b antibody
    • C4B_2 antibody
    • C4B1 antibody
    • C4B12 antibody
    • C4B2 antibody
    • C4B3 antibody
    • C4B5 antibody
    • C4BD antibody
    • C4d antibody
    • C4d fragment antibody
    • C4F antibody
    • C4S antibody
    • CH antibody
    • Chido form of C4 antibody
    • CO4 antibody
    • CO4A_HUMAN antibody
    • Complement C4 A antibody
    • Complement C4 B antibody
    • Complement C4 gamma chain antibody
    • complement C4-A antibody
    • complement C4-B antibody
    • complement C4-B-like antibody
    • complement C4B1a antibody
    • Complement component 4A antibody
    • complement component 4A (Rodgers blood group) antibody
    • Complement component 4B antibody
    • complement component 4B (Chido blood group) antibody
    • complement component 4B (Chido blood group), copy 2 antibody
    • complement component 4B (Chido blood group)-like antibody
    • Complement component 4F antibody
    • Complement component 4S antibody
    • CPAMD2 antibody
    • CPAMD3 antibody
    • DADB-112B14.11 antibody
    • MGC164979 antibody
    • RG antibody
    • Rodgers form of C4 antibody
    see all

画像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C4d antibody (ab87424)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C4d antibody (ab87424)
    IHC image of C4d staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab87424, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • Western blot - Anti-C4d antibody (ab87424)
    Western blot - Anti-C4d antibody (ab87424)
    Anti-C4d antibody (ab87424) at 1 µg/ml + Human Plasma Total Protein Lysate (ab83997) at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution (Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) )

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 193 kDa
    Observed band size: 193,85 kDa why is the actual band size different from the predicted?
    Additional bands at: 110 kDa, 130 kDa, 30 kDa, 48 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 30 seconds


    C4d circulates in blood as a disulfide-linked trimer of an alpha, beta and gamma chain. Our immunogen sequence is within the alpha chain. The band at 85kDa could correspond to the alpha chain.
  • Immunocytochemistry/ Immunofluorescence - Anti-C4d antibody (ab87424)
    Immunocytochemistry/ Immunofluorescence - Anti-C4d antibody (ab87424)

    ICC image of ab87424 stained HepG2 cells. The cells were 100% Methanol fixed (5 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 (ab87424, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit 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. This antibody also gave a positive result in 100% Methanol fixed (5 min) HeLa, and MCF-7 cells at 5µg/ml.

参考文献 (1)

ab87424 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。

ab87424 は 1 報の論文で使用されています。

  • Pérez-Alós L  et al. Combining MAP-1:CD35 or MAP-1:CD55 fusion proteins with pattern-recognition molecules as novel targeted modulators of the complement cascade. FASEB J 33:12723-12734 (2019). PubMed: 31469600

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

1-2 of 2 Abreviews or Q&A

Question

Dear Miss/Mr,
C4d is a split product derived from C4 and is generated after complemet activation. Complement activation only occurs extracelulary. Therefore C4d ONLY can be generated extracellulary and covalently deposited on cell surfaces. It is truth that hepatocytes produce C4, the native protein, but C4d only can be deposited on the SURFACE of cells. There is plenty of basic literature about it. My question is whether your antibody recognizes the native form of C4 or only the split product C4d (neoantigen). In the case of the antibody specifically recognizes C4d and not C4, is this molecule internalized? Thank you very much for all your help. Yours sincerely,

Read More
Answer

Thank you for your inquiry.

The immungens for these antibodies correspond to sequences within the C4d. This will also lead to the recognition of the complement precursor that is intra cellular since the complement precursor also contains theC4d sequence.

In a healthy sample there should be not complement deposited on cell surfaces. The lab did not test disease models and therefore we do not have an image of C4d that is deposited on cell surfaces.

We will update the on-line datasheet for this product as soon as we receive additional data.


We apologize for any inconvenience this may cause.

Read More

Question

Inquiry: Dear Miss/Mr, I am thinking about buying the anti-C4d antibodies (ab87424) and (ab64157) for immunohistochemistry. In these websites: https://www.abcam.com/C4d-antibody-ab64157.html#C4d-Primary-antibodies-ab64157-3.jpeg https://www.abcam.com/C4d-antibody-ab87424.html#C4d-Primary-antibodies-ab87424-3.jpg you show pictures of a hepatocarcinoma stained for C4d with both antibodies. However, as shown in the pictures, the staining is found mainly in the cytoplasm. C4d is complement fragment deposited on cell membrane after complement activation. My question is why C4d staining shown in the pictures is cytoplasmatic? Is an artefact? Is C4d internalized? On that case do you have any reference to confirm that C4d is internalized? Thank you very much for all your help. Yours sincerely,

Read More
Answer

Thank you for your inquiry.

The liver (mainly hepatocytes) is responsible for biosynthesis of 90% of plasma complement components and their soluble regulators, including the classical (C1r/s, C2, C4, C4bp), alternative (C3, factor B), lectin (MBL, MASP1-3, Map19), terminal (C5, C6, C8, C9) pathways of the complement system, and soluble regulators (factors I, H, and C1 inhibitor).

The images shown on the datasheets of ab87424 and ab64157 are of liver samples. Since these cell produce the complement they will show it in cytoplasma. The secreted complement will be washed away in IHC and ICC and therefore cannot be detected.

Please follow this link to review a publication about complement:

http://www.cmi.ustc.edu.cn/3/5/333.pdf

Complement is in the blood stream andis supposed to be deposited on bacteria or other invading microorganisms. In some disease like lupus etcthe tightly regulatedsystem gets disregulated and therefore complement deposits can be found on body cells. I suggest to review the available literature to get more information.

I hope this information is helpful and wish you good luck with your research.

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