Anti-C5/C5a Antibody Picoband™

Product Name

Anti-C5/C5a Antibody Picoband™

View all C5 Antibodies

SKU/Catalog Number

PB9187

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-C5/C5a Antibody Picoband™ catalog # PB9187. Tested in IHC, WB applications. This antibody reacts with Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-C5/C5a Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9187)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived rat C5a recombinant protein (Position: D1-R77). Rat C5a shares 62% and 82% amino acid (aa) sequences identity with human and mouse C5a, respectively.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PB9187 is reactive to C5 in Rat

Applications

PB9187 is guaranteed for IHC, WB Boster Guarantee

Observed Molecular Weight

115 kDa

Calculated molecular weight

188.305kDa

Background of C5

C5a is a protein fragment released from complement component C5. This gene is mapped to 9q33.2. The protein encoded by this gene is the fifth component of complement, which plays an important role in inflammatory and cell killing processes. This protein is comprised of alpha and beta polypeptide chains that are linked by a disulfide bridge. An activation peptide, C5a, which is an anaphylatoxin that possesses potent spasmogenic and chemotactic activity, is derived from the alpha polypeptide via cleavage with a convertase. C5a plays a key role in increasing migration and adherence of neutrophils and monocytes to vessel walls. Mutations in this gene cause complement component 5 deficiency, a disease where patients show a propensity for severe recurrent infections. Defects in this gene have also been linked to susceptibility to liver fibrosis and to rheumatoid arthritis.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Rat, By Heat
Western blot, 0.1-0.5μg/ml, Rat

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of C5A using anti-C5A antibody (PB9187).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: recombinant rat C5A protein 0.5 ng.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-C5A antigen affinity purified polyclonal antibody (Catalog # PB9187) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for C5A at approximately 37 kDa. The expected band size for C5A is at 37 kDa.

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Figure 2. Western blot analysis of C5A using anti-C5A antibody (PB9187).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat liver tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-C5A antigen affinity purified polyclonal antibody (Catalog # PB9187) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for C5A at approximately 115 kDa. The expected band size for C5A is at 115 kDa.

Click image to see more details

Figure 3. IHC analysis of C5A using anti-C5A antibody (PB9187).
C5A was detected in a paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in EDTAbuffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-C5A Antibody (PB9187) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Gene/Protein Information For C5 (Source: Uniprot.org, NCBI)

Alternative Names

C5; Complement Component C5/C5a C5 C5Da, C5b, CPAMD4, ECLZB, C5 complement C5 complement C5|C3 and PZP-like alpha-2-macroglobulin domain-containing protein 4|C5a anaphylatoxin|anaphylatoxin C5a analog|complement component 5|prepro-C5

For more info on C5, check out the C5 Infographic

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In this infographic, you will see the following information for C5: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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