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Knockout Tested Rabbit Recombinant Monoclonal CD33 antibody. Carrier free. Suitable for IP, WB and reacts with Human samples.

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Images

Immunoprecipitation - Anti-CD33 antibody [EPR4423] - Low endotoxin, Azide free (AB215383), expandable thumbnail
  • OI-RD Scanning - Anti-CD33 antibody [EPR4423] - Low endotoxin, Azide free (AB215383), expandable thumbnail
  • Western blot - Anti-CD33 antibody [EPR4423] - Low endotoxin, Azide free (AB215383), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWB
Human
Tested
Tested

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

2 products for Alternative Version

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: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

Knockout Tested Rabbit Recombinant Monoclonal CD33 antibody. Carrier free. Suitable for IP, WB and reacts with Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR4423
Purification technique
Affinity purification Protein A
Dissociation constant
1.76 x 10-11 M
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab215383 is the carrier-free version of Anti-CD33 antibody [EPR4423] ab134115.

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.

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.

Our Low endotoxin, azide-free formats have low endotoxin level (1 EU/mg, determined by the TAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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.

3 product images

  • Immunoprecipitation - Anti-CD33 antibody [EPR4423] - Low endotoxin, Azide free (ab215383), expandable thumbnail

    Immunoprecipitation - Anti-CD33 antibody [EPR4423] - Low endotoxin, Azide free (ab215383)

    This IP data was generated using the same anti-CD33 antibody clone, EPR4423, in a different buffer formulation (cat# Anti-CD33 antibody [EPR4423] ab134115).

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

    Predicted band size: 40 kDa

  • OI-RD Scanning - Anti-CD33 antibody [EPR4423] - Low endotoxin, Azide free (ab215383), expandable thumbnail

    OI-RD Scanning - Anti-CD33 antibody [EPR4423] - Low endotoxin, Azide free (ab215383)

    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] - Low endotoxin, Azide free (ab215383), expandable thumbnail

    Western blot - Anti-CD33 antibody [EPR4423] - Low endotoxin, Azide free (ab215383)

    False colour image of Western blot: Anti-CD33 antibody [EPR4423] 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, Anti-CD33 antibody [EPR4423] 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD33 antibody [EPR4423] ab134115).

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