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CD33 Recombinant Rabbit Monoclonal Antibody [PSH27-78]

Usd: 385

Specification

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Catalog# HA724723

CD33 Recombinant Rabbit Monoclonal Antibody [PSH27-78]

Application

  • WB

  • IHC-P

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

CD33 Recombinant Rabbit Monoclonal Antibody [PSH27-78]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human CD33 aa 1-282.

Species Reactivity

Human

Validated Applications

WB, IHC-P

Target Molecular Weight

Predicted band size: 40 kDa

Positive Control

TF-1 (Human erythroleukemia cell) cell lysate, THP-1 (Human acute monoblastic leukemia cell) cell lysate, HL-60 (Human acute promyelocytic leukemia cell) cell lysate, HEL (Human erythroleukemia cell) cell lysate, RPMI8226 (Human multiple myeloma cell) cell lysate.

Conjugation

unconjugated

Clone Number

PSH27-78

Reactivity Data

Tested Tested Tested
Verified (internally validated)
Published Tested
Reported in literature (not internally validated)
Predicted Tested
Predicted reactive (based on sequence homology)
Not recommended Tested
Not recommended (failed internal validation)
WB IHC-P FC IF-Cell
human
Tested Tested
Tested Tested
Not Recommended
Not Recommended

Product Features

Form

Liquid

Concentration

0.5 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:500

Target

Function

CD33 or Siglec-3 (sialic acid binding Ig-like lectin 3, SIGLEC3, SIGLEC-3, gp67, p67) is a transmembrane receptor expressed on cells of myeloid lineage. It is usually considered myeloid-specific, but it can also be found on some lymphoid cells. It binds sialic acids, therefore is a member of the SIGLEC family of lectins. CD33 can be stimulated by any molecule with sialic acid residues such as glycoproteins or glycolipids. Upon binding, the immunoreceptor tyrosine-based inhibition motif (ITIM) of CD33, present on the cytosolic portion of the protein, is phosphorylated and acts as a docking site for Src homology 2 (SH2) domain-containing proteins like SHP phosphatases. This results in a cascade that inhibits phagocytosis in the cell.

Background References

1. Albinger N et al. Primary CD33-targeting CAR-NK cells for the treatment of acute myeloid leukemia. Blood Cancer J. 2022 Apr

2. Willier S et al. CLEC12A and CD33 coexpression as a preferential target for pediatric AML combinatorial immunotherapy. Blood. 2021 Feb

Subcellular Location

Cell membrane; Peroxisome.

UNIPROT

SwissProt: P20138 Human

Synonyms

CD 33 antibody

CD33 antibody

CD33 antigen (gp67) antibody

CD33 antigen antibody

CD33 molecule antibody

CD33_HUMAN antibody

FLJ00391 antibody

gp67 antibody

My9 antibody

Myeloid cell surface antigen CD33 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE),Independent antibody validation (IAV)</span><br /><br />Western blot analysis of CD33 on different lysates with Rabbit anti-CD33 antibody (<a href="/products/HA724723" style="font-weight: bold;text-decoration: underline;">HA724723</a>) at 1/2,000 dilution. <br /> <br />Lane 1: TF-1 (Human erythroleukemia cell) cell lysate <br />Lane 2: Jurkat (Human T-lymphoblastic cells) cell lysate <br />Lane 3: THP-1 (Human acute monoblastic leukemia cell) cell lysate <br />Lane 4: HL-60 (Human acute promyelocytic leukemia cell) cell lysate <br /> <br />Lysates/proteins at 15 µg/Lane. <br />Exposure time: 20 seconds; ECL: K1801 <br /> <br />Negative Control: Jurkat (PMID: 30519686). <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724723" style="font-weight: bold;text-decoration: underline;">HA724723</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 40 kDa <br />Observed band size: 67-75 kDa

    ☑ Relative expression (RE),Independent antibody validation (IAV)

    Western blot analysis of CD33 on different lysates with Rabbit anti-CD33 antibody (HA724723) at 1/2,000 dilution.

    Lane 1: TF-1 (Human erythroleukemia cell) cell lysate
    Lane 2: Jurkat (Human T-lymphoblastic cells) cell lysate
    Lane 3: THP-1 (Human acute monoblastic leukemia cell) cell lysate
    Lane 4: HL-60 (Human acute promyelocytic leukemia cell) cell lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 20 seconds; ECL: K1801

    Negative Control: Jurkat (PMID: 30519686).

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724723, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 40 kDa
    Observed band size: 67-75 kDa

  • <span style="font-weight: bold;">☑ Independent antibody validation (IAV)</span><br /><br />Western blot analysis of CD33 on different lysates with Rabbit anti-CD33 antibody (<a href="/products/HA724723" style="font-weight: bold;text-decoration: underline;">HA724723</a>) at 1/2,000 dilution. <br /> <br />Lane 1: HEL (Human erythroleukemia cell) cell lysate <br />Lane 2: RPMI8226 (Human multiple myeloma cell) cell lysate <br /> <br />Lysates/proteins at 15 µg/Lane. <br />Exposure time: 20 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724723" style="font-weight: bold;text-decoration: underline;">HA724723</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 40 kDa <br />Observed band size: 67-75 kDa

    ☑ Independent antibody validation (IAV)

    Western blot analysis of CD33 on different lysates with Rabbit anti-CD33 antibody (HA724723) at 1/2,000 dilution.

    Lane 1: HEL (Human erythroleukemia cell) cell lysate
    Lane 2: RPMI8226 (Human multiple myeloma cell) cell lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 20 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724723, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 40 kDa
    Observed band size: 67-75 kDa

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Liver <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724723" style="font-weight: bold;text-decoration: underline;">HA724723</a>, 1/500, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Liver
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724723, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Placenta <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724723" style="font-weight: bold;text-decoration: underline;">HA724723</a>, 1/500, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Placenta
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724723, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Tonsil <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724723" style="font-weight: bold;text-decoration: underline;">HA724723</a>, 1/500, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Tonsil
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724723, 1/500, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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