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Knockout Tested Rabbit Recombinant Monoclonal CD33 antibody. Suitable for IP, Flow Cyt, WB and reacts with Human samples. Cited in 3 publications.

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Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% 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
IPFlow CytWBIHC-P
Human
Tested
Tested
Tested
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

1/10 - 1/100

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000 - 1/10000

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse

Dilution info

-

Notes

-

Target data

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:15597323, PubMed:11320212). 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:28325905, PubMed:10887109). These phosphorylations provide docking sites for the recruitment and activation of protein-tyrosine phosphatases PTPN6/SHP-1 and PTPN11/SHP-2 (PubMed:10556798, PubMed:10206955, 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

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Knockout Tested Rabbit Recombinant Monoclonal CD33 antibody. Suitable for IP, Flow Cyt, WB and reacts with Human samples. Cited in 3 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Clone number

EPR4423

Purification technique

Affinity purification Protein A

Dissociation constant

1.76 x 10-11 M

Concentration
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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

Stable for 12 months at -20°C

Notes

Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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.

6 product images

  • Flow Cytometry - Anti-CD33 antibody [EPR4423] (ab134115), expandable thumbnail

    Flow Cytometry - Anti-CD33 antibody [EPR4423] (ab134115)

    Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte, Left) / THP1 (Human monocytic leukemia monocyte, Right) labeling CD33 with ab134115 at 1/500 dilution (red). Goat anti rabbit IgG (Alexa Fluor® 647, ab150079) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as isotype control (Black). Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue).

    Negative control: Jurkat.
    Gated on viable cells.

  • Immunoprecipitation - Anti-CD33 antibody [EPR4423] (ab134115), expandable thumbnail

    Immunoprecipitation - Anti-CD33 antibody [EPR4423] (ab134115)

    ab134115 (purified) at 1/60 immunoprecipitating CD33 in 10 μg THP-1 (Lanes 1 and 2, observed at 67-75 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

    All lanes: Immunoprecipitation - Anti-CD33 antibody [EPR4423] (AB134115)

    Predicted band size: 40 kDa

  • Western blot - Anti-CD33 antibody [EPR4423] (ab134115), expandable thumbnail

    Western blot - Anti-CD33 antibody [EPR4423] (ab134115)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

    All lanes: Western blot - Anti-CD33 antibody [EPR4423] (AB134115) at 1/5000 dilution

    Lane 1: THP-1 cell lysate at 10 µg

    Lane 2: U937 cell lysate at 10 µg

    Secondary

    All lanes: HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size: 40 kDa

    Observed band size: 67 kDa, 75 kDa

  • OI-RD Scanning - Anti-CD33 antibody [EPR4423] (ab134115), expandable thumbnail

    OI-RD Scanning - Anti-CD33 antibody [EPR4423] (ab134115)

    We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
    Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

  • Western blot - Anti-CD33 antibody [EPR4423] (ab134115), expandable thumbnail

    Western blot - Anti-CD33 antibody [EPR4423] (ab134115)

    All lanes: Western blot - Anti-CD33 antibody [EPR4423] (AB134115) at 1/1000 dilution

    Lane 1: THP1 cell lysate at 10 µg

    Lane 2: U937 cell lysate at 10 µg

    Lane 3: HL60 cell lysate at 10 µg

    Lane 4: HepG2 cell lysate at 10 µg

    Lane 5: HeLa cell lysate at 10 µg

    Secondary

    All lanes: Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

    Predicted band size: 40 kDa

    Observed band size: 67-75 kDa

  • Western blot - Anti-CD33 antibody [EPR4423] (ab134115), expandable thumbnail

    Western blot - Anti-CD33 antibody [EPR4423] (ab134115)

    False colour image of Western blot: Anti-CD33 antibody [EPR4423] 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, ab134115 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 [EPR4423] (AB134115) at 1/1000 dilution

    Lane 1: Wild-type 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

    Observed band size: 60-80 kDa

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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