Recombinant Anti-CD33 antibody [EPR23051-101] (ab269456)
![Western blot - Anti-CD33 antibody [EPR23051-101] (ab269456)](https://www.abcam.com/ps/products/269/ab269456/Images/ab269456-513047-anti-cd33-antibody-epr23051-101-western-blot-wildtype-thp1-cd33-knockout-thp1-hl60-jurkat.jpg "Western blot - Anti-CD33 antibody [EPR23051-101] (ab269456)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23051-101] to CD33
- Suitable for: IHC-P, ICC/IF, IP, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD33 antibody [EPR23051-101]
See all CD33 primary antibodies -
Description
Rabbit monoclonal [EPR23051-101] to CD33 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, ICC/IF, IP, WBmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: U937 and THP-1 whole cell lysates; Human liver lysate; Wild-type THP-1 and HL-60 cell lysates. IHC-P: Human liver, Human Hodgkiin's lymphoma and Human clear cell renal cell carcinoma tissue . ICC/IF: THP-1 cells. IP: THP-1 cells.
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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, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23051-101 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab269456 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 |
|---|---|---|
| IHC-P | (1) |
1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
| ICC/IF | (1) |
1/50.
|
| IP |
1/30.
|
|
| WB | (1) |
1/1000. Predicted molecular weight: 40 kDa.
|
| Notes |
|---|
|
IHC-P
1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
ICC/IF
1/50. |
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IP
1/30. |
|
WB
1/1000. 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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Western blot - Anti-CD33 antibody [EPR23051-101] (ab269456)All lanes : Anti-CD33 antibody [EPR23051-101] (ab269456) 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.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
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?False colour image of Western blot: Anti-CD33 antibody [EPR23051-101] 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, ab269456 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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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR23051-101] (ab269456)](https://www.abcam.com/ps/products/269/ab269456/Images/ab269456-3-anti-cd33-antibody-epr23051-101-immunohistochemistry-hodgkins-lymphoma-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR23051-101] (ab269456)Immunohistochemical analysis of paraffin-embedded Human Hodgkin's lymphoma tissue labeling CD33 with ab269456 at 1/200 dilution (2.96 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on human Hodgkin's lymphoma.
The section was incubated with ab269456 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 solution 2) for 20 mins.
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![Immunoprecipitation - Anti-CD33 antibody [EPR23051-101] (ab269456)](https://www.abcam.com/ps/products/269/ab269456/Images/ab269456-9-anti-cd33-antibody-epr23051-101-immunoprecipitation-thp1-human.jpg)
Immunoprecipitation - Anti-CD33 antibody [EPR23051-101] (ab269456)CD33 was immunoprecipitated from 0.35 mg THP-1 (Human monocytic leukemia monocyte) whole cell lysate with ab269456 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab269456 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: THP-1 whole cell lysate 10ug.
Lane 2: ab269456 IP in THP-1 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab269456 in THP-1 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 23 seconds.
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![Immunocytochemistry/ Immunofluorescence - Anti-CD33 antibody [EPR23051-101] (ab269456)](https://www.abcam.com/ps/products/269/ab269456/Images/ab269456-6-anti-cd33-antibody-epr23051-101-immunocytochemistry-thp1-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CD33 antibody [EPR23051-101] (ab269456)Immunofluorescent analysis of 100% methanol-fixed THP-1 (human monocytic leukemia monocyte) and Jurkat (human T cell leukemia T lymphocyte) cells labeling CD33 with ab26946 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). Confocal image showing membranous staining in THP-1 cells. The nuclear counter stain is DAPI (Blue). Tubilin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (Red).
Control: Used PBS instead of primary antibody, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.Negative control: Jurkat (PMID: 30519686).
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![Western blot - Anti-CD33 antibody [EPR23051-101] (ab269456)](https://www.abcam.com/ps/products/269/ab269456/Images/ab269456-8-anti-cd33-antibody-epr23051-101-western-blot-human.jpg)
Western blot - Anti-CD33 antibody [EPR23051-101] (ab269456)All lanes : Anti-CD33 antibody [EPR23051-101] (ab269456) at 1/1000 dilution
Lane 1 : U937 (human histiocytic lymphoma monocyte), whole cell lysate
Lane 2 : THP-1 (human monocytic leukemia monocyte), whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 40 kDa
Observed band size: 67-75 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 70 seconds.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:16380601, 30519686).
Negative Control:Jurkat (PMID:30519686).
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![Western blot - Anti-CD33 antibody [EPR23051-101] (ab269456)](https://www.abcam.com/ps/products/269/ab269456/Images/ab269456-7-anti-cd33-antibody-epr23051-101-western-blot-human-2.jpg)
Western blot - Anti-CD33 antibody [EPR23051-101] (ab269456)All lanes : Anti-CD33 antibody [EPR23051-101] (ab269456) at 1/1000 dilution
Lane 1 : Human liver tissue lysate
Lane 2 : Human skeletal muscle tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 40 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 70 seconds.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:16380601, 30519686).
Negative control:Human skeletal muscle.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR23051-101] (ab269456)](https://www.abcam.com/ps/products/269/ab269456/Images/ab269456-4-anti-cd33-antibody-epr23051-101-immunohistochemistry-clear-cell-renal-cell-carcinoma-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR23051-101] (ab269456)Immunohistochemical analysis of paraffin-embedded Human clear cell renal cell carcinoma tissue labeling CD33 with ab269456 at 1/200 dilution (2.96 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on infiltrated myeloid cells of human clear cell renal cell carcinoma The section was incubated with ab269456 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 solution 2) for 20 mins.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR23051-101] (ab269456)](https://www.abcam.com/ps/products/269/ab269456/Images/ab269456-2-anti-cd33-antibody-epr23051-101-immunohistochemistry-liver-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR23051-101] (ab269456)Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling CD33 with ab269456 at 1/200 dilution (2.96 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on mainly Kuffer cells of human liver (PMID:25721896). The section was incubated with ab269456 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 solution 2) for 20 mins.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR23051-101] (ab269456)](https://www.abcam.com/ps/products/269/ab269456/Images/ab269456-5-anti-cd33-antibody-epr23051-101-immunohistochemistry-skeletal-muscle-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR23051-101] (ab269456)Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling CD33 with ab269456 at 1/200 dilution (2.96 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Negative control：Human skeletal muscle.
The section was incubated with ab269456 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
-
![Anti-CD33 antibody [EPR23051-101] (ab269456)](https://www.abcam.com/ps/products/269/ab269456/Images/ab269456-10-benefits-of-recombinant-antibodies.png)
Anti-CD33 antibody [EPR23051-101] (ab269456)
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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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab269456 has been referenced in 2 publications.
- Nian Z et al. Rapamycin Pretreatment Rescues the Bone Marrow AML Cell Elimination Capacity of CAR-T Cells. Clin Cancer Res 27:6026-6038 (2021). PubMed: 34233960
- Rong X et al. PET/CT Imaging of Activated Cancer-Associated Fibroblasts Predict Response to PD-1 Blockade in Gastric Cancer Patients. Front Oncol 11:802257 (2021). PubMed: 35155199