HRP Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab195198)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- HRP Rabbit monoclonal [EPR3633(2)] to Calnexin - ER Membrane Marker
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
- Conjugation: HRP
Overview
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Product name
HRP Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker
See all Calnexin primary antibodies -
Description
HRP Rabbit monoclonal [EPR3633(2)] to Calnexin - ER Membrane Marker -
Host species
Rabbit -
Conjugation
HRP -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HepG2, A431 and HeLa whole cell lysates. IHC-P: normal human colon 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. Avoid freeze / thaw cycle. Store In the Dark. -
Storage buffer
pH: 7.40
Preservative: 0.1% 10% Proclin 300 Solution
Constituents: 30% Glycerol (glycerin, glycerine), 1% BSA, PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3633(2) -
Isotype
IgG -
Research areas
Images
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All lanes : HRP Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab195198) at 1/5000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : CANX (Calnexin) knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 68 kDaab195198 was shown to recognize Calnexin in wild-type HAP1 cells as signal was lost at the expected MW in CANX (Calnexin) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CANX (Calnexin) knockout samples were subjected to SDS-PAGE. Ab195198 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.
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IHC image of Calnexin staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab195198 at 1/500 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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All lanes : HRP Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab195198) at 1/5000 dilution
Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 :HeLa whole cell lysate (ab150035)
Lysates/proteins at 10 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 82 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutesThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab195198 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
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