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  1. Link

    c5--c5b-antibody-epr23940-3-ab275931.pdf

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Immunology Innate Immunity Macrophage / Inflamm.
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RecombinantRabMAb

Recombinant Anti-C5 / C5b antibody [EPR23940-3] (ab275931)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
  • Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
  • Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
  • Anti-C5 / C5b antibody [EPR23940-3] (ab275931)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23940-3] to C5 / C5b
  • Suitable for: WB, IHC-P
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-C5 / C5b antibody [EPR23940-3]
    See all C5 / C5b primary antibodies
  • Description

    Rabbit monoclonal [EPR23940-3] to C5 / C5b
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human plasma and liver tissue lysate; Mouse plasma; Rat plasma. IHC-P: Human liver and breast tissue.
  • General notes

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR23940-3
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Macrophage / Inflamm.
    • Immunology
    • Innate Immunity
    • Complement
    • MAC
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress

Associated products

  • Alternative Versions

    • Anti-C5 / C5b antibody [EPR23940-3] - BSA and Azide free (ab275933)
  • Compatible Secondaries

    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab275931 in the following tested applications.

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

Application Abreviews Notes
WB
1/1000. Detects a band of approximately 70, 190 kDa (predicted molecular weight: 188 kDa).
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
WB
1/1000. Detects a band of approximately 70, 190 kDa (predicted molecular weight: 188 kDa).
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Application notes
Is unsuitable for Flow Cyt,ICC or IP.

Target

  • Relevance

    C5 is synthesised in the liver as a single polypeptide chain. Before secretion the molecule is glycosylated and secreted into plasma as a 190kDa glycoprotein consisting of a disulphide linked alpha-chain (115kDa) and beta-chain (75kDa). C5 precursor is first processed by the removal of 4 basic residues, forming two chains, beta and alpha, linked by a disulfide bond. C5 convertase activates C5 by cleaving the alpha chain, releasing C5a anaphylatoxin and generating C5b (beta chain + alpha' chain).
  • Cellular localization

    Secreted
  • Database links

    • Entrez Gene: 727 Human
    • Entrez Gene: 362119 Rat
    • Omim: 120900 Human
    • SwissProt: P01031 Human
    • SwissProt: P06684 Mouse
    • SwissProt: P08650 Rat
    • Alternative names

      • C5 antibody
      • Complement C5 antibody
      • Complement component 5 antibody
      • CPAMD4 antibody

    Images

    • Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
      Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
      All lanes : Anti-C5 / C5b antibody [EPR23940-3] (ab275931) at 1/1000 dilution

      Lane 1 : HepG2 cell lysate
      Lane 2 : U-2 OS cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 188 kDa
      Observed band size: 165-180 kDa why is the actual band size different from the predicted?



      False colour image of Western blot: Anti-C5 / C5b antibody [EPR23940-3] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab275931 was shown to bind specifically to C5 / C5b. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)

      Immunohistochemical analysis of paraffin-embedded human breast tissue labeling C5/C5b with ab275931 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on plasma of human breast. The section was incubated with ab275931 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
      Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
      All lanes : Anti-C5 / C5b antibody [EPR23940-3] (ab275931) at 1/1000 dilution

      Lane 1 : Mouse plasma
      Lane 2 : Rat plasma

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

      Predicted band size: 188 kDa
      Observed band size: 70 kDa why is the actual band size different from the predicted?



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

      The molecular weight observed is consistent with what has been described in the literature (PMID: 23401592, 25387724).

      Exposure time: 48 seconds.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)

      Immunohistochemical analysis of paraffin-embedded human liver tissue labeling C5/C5b with ab275931 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human liver. The section was incubated with ab275931 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
      Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
      All lanes : Anti-C5 / C5b antibody [EPR23940-3] (ab275931) at 1/1000 dilution

      Lane 1 : Human plasma at 10 µg
      Lane 2 : Human liver tissue lysate at 40 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 188 kDa
      Observed band size: 190,70 kDa why is the actual band size different from the predicted?



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

      Samples are non-boiled as boiling may cause protein aggregates. 

      The molecular weight observed is consistent with what has been described in the literature (PMID: 24071736, 23401592, 25387724)

      Exposure time: 59 seconds.

       

    • Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
      Anti-C5 / C5b antibody [EPR23940-3] (ab275931)

    Protocols

    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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab275931? Please let us know so that we can cite the reference in this datasheet.

    ab275931 has not yet been referenced specifically in any publications.

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