Anti-CD33 antibody [EPR24370-124] KO tested (ab270942)

Rabbit Recombinant Monoclonal CD33 antibody. Suitable for WB, IHC-P and reacts with Human samples.

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Images

Western blot - Anti-CD33 antibody [EPR24370-124] (AB270942), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	Flow Cyt	WB	IHC-P
Human	Not recommended	Not recommended	Not recommended	Tested	Tested
Mouse	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info -	Notes -
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target data

See full target information CD33

Function

Sialic-acid-binding immunoglobulin-like lectin (Siglec) that plays a role in mediating cell-cell interactions and in maintaining immune cells in a resting state (PubMed:10611343, PubMed:11320212, PubMed:15597323). Preferentially recognizes and binds alpha-2,3- and more avidly alpha-2,6-linked sialic acid-bearing glycans (PubMed:7718872). Upon engagement of ligands such as C1q or syalylated glycoproteins, two immunoreceptor tyrosine-based inhibitory motifs (ITIMs) located in CD33 cytoplasmic tail are phosphorylated by Src-like kinases such as LCK (PubMed:10887109, PubMed:28325905). These phosphorylations provide docking sites for the recruitment and activation of protein-tyrosine phosphatases PTPN6/SHP-1 and PTPN11/SHP-2 (PubMed:10206955, PubMed:10556798, PubMed:10887109). In turn, these phosphatases regulate downstream pathways through dephosphorylation of signaling molecules (PubMed:10206955, PubMed:10887109). One of the repressive effect of CD33 on monocyte activation requires phosphoinositide 3-kinase/PI3K (PubMed:15597323).

Alternative names

CD33, SIGLEC3, Myeloid cell surface antigen CD33, Sialic acid-binding Ig-like lectin 3, gp67, Siglec-3

Recommended products

Rabbit Recombinant Monoclonal CD33 antibody. Suitable for WB, IHC-P and reacts with Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR24370-124
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Please Note: Clone [EPR24370-124] (this product) is different to that of ab199432 [SP266] for the same target.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

Western blot - Anti-CD33 antibody [EPR24370-124] (ab270942)
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.
All lanes: Western blot - Anti-CD33 antibody [EPR24370-124] (ab270942) at 1/1000 dilution
Lane 1: THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg
Lane 2: Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 3: HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg
Lane 4: TF-1 (human erythroleukemia erythroblast) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 40 kDa
Observed band size: 67-75 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR24370-124] (ab270942)
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR24370-124] (ab270942)
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR24370-124] (ab270942)
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR24370-124] (ab270942)
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR24370-124] (ab270942)
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD33 antibody [EPR24370-124] (ab270942)
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.
Western blot - Anti-CD33 antibody [EPR24370-124] (ab270942)
False colour image of Western blot: Anti-CD33 antibody [EPR24370-124] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control 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.
All lanes: Western blot - Anti-CD33 antibody [EPR24370-124] (ab270942) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 10 µg
Lane 2: Western blot - Human CD33 knockout THP-1 cell lysate (ab273785)
Lane 2: CD33 knockout THP-1 cell lysate at 10 µg
Lane 3: HL-60 cell lysate at 10 µg
Lane 4: Jurkat cell lysate at 10 µg
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