Anti-CD9 antibody [EPR2949] (ab92726)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2949] to CD9
- Suitable for: WB, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-CD9 antibody [EPR2949]
See all CD9 primary antibodies -
Description
Rabbit monoclonal [EPR2949] to CD9 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P RatHumanIP HumanWB MouseHuman -
Immunogen
Synthetic peptide within Human CD9 aa 200 to the C-terminus. The exact sequence is proprietary.
Database link: P21926 -
Positive control
- WB: HeLa, HuT-78, MCF7 and U87-MG cell lysates. Mouse heart and kidney lysate; Rat brain lysate. IHC-P: Human papillary carcinoma , astrocytoma, brain, kidney and tonsil tissue; Rat spleen tissue. IP: HeLa whole cell lysate (ab150035).
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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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2949 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-CD9 antibody [EPR2949] (ab92726) at 1/1000 dilution
Lane 1 : Wild-type A549 whole cell lysate
Lane 2 : CD9 knockout A549 whole cell lysate
Lane 3 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 25 kDaLanes 1 - 3: Merged signal (red and green). Green - ab92726 observed at 22 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab92726 was shown to recognize CD9 in wild-type A549 cells as signal was lost at the expected MW in CD9 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CD9 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab92726 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemical staining of paraffin embedded rat spleen with purified ab92726 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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ab92726 (purified) at 1/20 immunoprecipitating CD9 in 10 μg HeLa (Lanes 1 and 2, observed at 24 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST
Dilution buffer and concentration: 5% NFDM/TBST
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Formalin-fixed, paraffin-embedded Papillary carcinoma of thyroid gland tissue stained for CD9 using ab92726 at 1/500 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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All lanes : Anti-CD9 antibody [EPR2949] (ab92726) at 1/1000 dilution
Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell). Whole cell lysates with 5% NFDM/TBST
Lane 2 : Daudi (Human Burkitt's lymphoma lymphoblast). Whole cell lysates. (negative/low expression control) with 5% NFDM/TBST
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 25 kDa
Observed band size: 22 kDa why is the actual band size different from the predicted?
Exposure time: 3 seconds
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Formalin-fixed, paraffin-embedded Normal kidney tissue stained for CD9 using ab92726 at 1/500 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Formalin-fixed, paraffin-embedded Astrocytoma tissue stained for CD9 using ab92726 at 1/500 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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All lanes : Anti-CD9 antibody [EPR2949] (ab92726) at 1/10000 dilution (purified)
Lane 1 : HeLa cell lysate
Lane 2 : HuT-78 cell lysate
Lane 3 : U87-MG cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 25 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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All lanes : Anti-CD9 antibody [EPR2949] (ab92726) at 1/2000 dilution (purified)
Lane 1 : mouse heart lysate
Lane 2 : mouse kidney lysate
Lane 3 : rat brain lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 25 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Formalin-fixed, paraffin-embedded Normal brain tissue stained for CD9 using ab92726 at 1/500 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemical staining of paraffin embedded human tonsil with purified ab92726 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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