Anti-Serpin A5 antibody [EPR23337-69] (ab270450)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23337-69] to Serpin A5
- Suitable for: WB, IP, IHC-P
- Reacts with: Human
Overview
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Product name
Anti-Serpin A5 antibody [EPR23337-69]
See all Serpin A5 primary antibodies -
Description
Rabbit monoclonal [EPR23337-69] to Serpin A5 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanIP HumanWB Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human serum, plasma and testis lysates; MCF7, HepG2 and MDA-MB-231 whole cell lysates. IHC-P: Human kidney and testis tissue. IP: Human serum lysate.
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23337-69 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-Serpin A5 antibody [EPR23337-69] (ab270450) at 1/1000 dilution
Lane 1 : Human serum lysate
Lane 2 : Human plasma lysate
Lane 3 : Human testis tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 45,52,80-90 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/molecular weight observed is consistent with what has been described in the literature. Bands detected between 80 kDa and 120 kDa could be either the dimeric form or serpin A5 in complex with a serine protease such as urokinase or plasma kallikrein. Bands around 50 kDa represent the intact and cleaved forms (PMID: 22206708).
Exposure time: 8 seconds.
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Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Serpin A5 with ab270450 at 1/5000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the human testis (PMID: 1372913). The section was incubated with ab270450 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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Serpin A5 was immunoprecipitated from 0.35 mg Human serum lysate with ab270450 at 1/30 dilution . Western blot was performed on the immunoprecipitate using ab270450 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Human serum lysate 10 ug
Lane 2: ab270450 IP in Human serum lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab270450 in Human serum lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
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All lanes : Anti-Serpin A5 antibody [EPR23337-69] (ab270450) at 1/1000 dilution
Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution
Predicted band size: 46 kDa
Observed band size: 45,52,80-90 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/molecular weight observed is consistent with what has been described in the literature. The band detected around 100 kDa could be either the dimeric form or serpin A5 in complex with a serine protease such as urokinase or kallikrein. Bands around 50 kDa represent the intact and cleaved forms (PMID: 22206708).
Exposure time: Lane 1: 3 minutes; Lane 2: 37 seconds; Lane 3: 3 minutes.
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Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Serpin A5 with ab270450 at 1/5000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the proximal tubular epithelium of the human kidney (PMID: 14696115). The section was incubated with ab270450 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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