Recombinant Anti-CD33 antibody [EPR24370-124] - BSA and Azide free (ab281568)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24370-124] to CD33 - BSA and Azide free
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD33 antibody [EPR24370-124] - BSA and Azide free
See all CD33 primary antibodies -
Description
Rabbit monoclonal [EPR24370-124] to CD33 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: THP-1, HL-60, TF-1 lysates. IHC-P: Human colon, liver, tonsil, Hodgkin's lymphoma and clear renal carcinoma tissues.
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General notes
Please Note: Clone [EPR24370-124] (this product) is different to that of ab199432 [SP266] for the same target.
ab281568 is the carrier-free version of ab270942.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. -
Storage buffer
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24370-124 -
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 ab281568 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 |
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IHC-P | (2) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa.
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Notes |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa. |
Target
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Function
Putative adhesion molecule of myelomonocytic-derived cells that mediates sialic-acid dependent binding to cells. Preferentially binds to alpha-2,6-linked sialic acid. The sialic acid recognition site may be masked by cis interactions with sialic acids on the same cell surface. In the immune response, may act as an inhibitory receptor upon ligand induced tyrosine phosphorylation by recruiting cytoplasmic phosphatase(s) via their SH2 domain(s) that block signal transduction through dephosphorylation of signaling molecules. Induces apoptosis in acute myeloid leukemia (in vitro). -
Tissue specificity
Monocytic/myeloid lineage cells. -
Sequence similarities
Belongs to the immunoglobulin superfamily. SIGLEC (sialic acid binding Ig-like lectin) family.
Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
Domain
Contains 2 copies of a cytoplasmic motif that is referred to as the immunoreceptor tyrosine-based inhibitor motif (ITIM). This motif is involved in modulation of cellular responses. The phosphorylated ITIM motif can bind the SH2 domain of several SH2-containing phosphatases. -
Post-translational
modificationsPhosphorylation of Tyr-340 is involved in binding to PTPN6 and PTPN11. Phosphorylation of Tyr-358 is involved in binding to PTPN6. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 945 Human
- Omim: 159590 Human
- SwissProt: P20138 Human
- Unigene: 83731 Human
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Alternative names
- CD 33 antibody
- CD33 antibody
- CD33 antigen (gp67) antibody
see all
Images
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All lanes : Anti-CD33 antibody [EPR24370-124] (ab270942) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : CD33 knockout THP-1 cell lysate
Lane 3 : HL-60 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 60-80 kDa why is the actual band size different from the predicted?This data was developed using ab270942, the same antibody clone in a different buffer formulation.
False colour image of Western blot: Anti-CD33 antibody [EPR24370-124] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab270942 was shown to bind specifically to CD33. A band was observed at 60-80 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CD33 knockout cell line ab273831 (knockout cell lysate ab273785). To generate this image, wild-type and CD33 knockout THP-1 cell lysates were analysed. 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-CD33 antibody [EPR24370-124] (ab270942) at 1/1000 dilution
Lane 1 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 2 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 3 : HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate
Lane 4 : TF-1 (human erythroleukemia erythroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 40 kDa
Observed band size: 67-75 kDa why is the actual band size different from the predicted?This data was developed using ab270942, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression molecular weight observed is consistent with what has been described in the literature (PMID:16380601, 30519686).
Negative Control: Jurkat (PMID:30519686).
Exposure time: Lanes 1-3:136 seconds; Lane 4:26 seconds.
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This data was developed using ab270942, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining on immune cells of human colon (PMID: 31462392). The section was incubated with ab270942 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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This data was developed using ab270942, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining on Kupffer cells of human liver (PMID:25721896). The section was incubated with ab270942 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
This data was developed using ab270942, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining in human tonsil. The section was incubated with ab270942 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
This data was developed using ab270942, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human Hodgkin's lymphoma tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining in human Hodgkin's lymphoma. The section was incubated with ab270942 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
This data was developed using ab270942, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human clear cell renal cell carcinoma tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining on scattered cells of human clear cell renal cell carcinoma. The section was incubated with ab270942 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
This data was developed using ab270942, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling CD33 with ab270942 at 1/1000 dilution (0.472 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining in human skeletal muscle. The section was incubated with ab270942 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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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Datasheet download
Certificate of Compliance
References (0)
ab281568 has not yet been referenced specifically in any publications.