Recombinant Anti-C5 / C5b antibody [EPR23940-3] (ab275931)
![Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)](https://www.abcam.com/ps/products/275/ab275931/Images/ab275931-480337-anti-c5-c5b-antibody-epr23940-3-western-blot-hepg2-u2-os.jpg "Western blot - 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
Related conjugates and formulations
Overview
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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.
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Positive control
- WB: Human plasma and liver tissue lysate; Mouse plasma; Rat plasma. IHC-P: Human liver and breast tissue.
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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
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
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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
Associated products
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Alternative Versions
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Compatible Secondaries
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).
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| IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Notes |
|---|
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WB
1/1000. Detects a band of approximately 70, 190 kDa (predicted molecular weight: 188 kDa). |
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IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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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
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Alternative names
- C5 antibody
- Complement C5 antibody
- Complement component 5 antibody
- CPAMD4 antibody
Images
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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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)](https://www.abcam.com/ps/products/275/ab275931/Images/ab275931-2-anti-c5c5b-antibody-epr239403-immunohistochemistry-human-breast.jpg)
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.
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![Western blot - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)](https://www.abcam.com/ps/products/275/ab275931/Images/ab275931-412151-anti-c5c5b-antibody-epr239403-western-blot-ug1.jpg)
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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-C5 / C5b antibody [EPR23940-3] (ab275931)](https://www.abcam.com/ps/products/275/ab275931/Images/ab275931-1-anti-c5c5b-antibody-epr239403-immunohistochemistry-human-liver.jpg)
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)](https://www.abcam.com/ps/products/275/ab275931/Images/ab275931-412150-anti-c5c5b-antibody-epr239403-western-blot-ug.jpg)
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.
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![Anti-C5 / C5b antibody [EPR23940-3] (ab275931)](https://www.abcam.com/ps/products/275/ab275931/Images/ab275931-5-benefits-of-recombinant-antibodies.png)
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab275931 has not yet been referenced specifically in any publications.