Name: Anti-CD33 antibody [EPR23051-101]
Brand: RabMAb
SKU: ab269456
Rating: 5 (3 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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
Reacts with: Human

Carrier Free

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.
Positive control
- WB: U937 and THP-1 whole cell lysates; Human liver lysate. 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.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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

Alternative Versions
- Anti-CD33 antibody [EPR23051-101] - BSA and Azide free (ab269461)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Anti-CD33 antibody [EPR23052-5] (ab264558)

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		1/1000. 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.
IP 1/30.
WB 1/1000. Predicted molecular weight: 40 kDa.

Application notes

Is unsuitable for Flow Cyt.

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
modifications

Phosphorylation 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.
Target information above from: UniProt accession P20138 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 945 Human
- Omim: 159590 Human
- SwissProt: P20138 Human
- Unigene: 83731 Human
Alternative names
- 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
- Myeloid cell surface antigen CD33 precursor antibody
- Myeloid differentiation antigen CD33 antibody
- p67 antibody
- Sialic acid binding Ig like lectin 3 antibody
- Sialic acid binding immunoglobulin like lectin 3 antibody
- Sialic acid-binding Ig-like lectin 3 antibody
- SIGLEC 3 antibody
- Siglec-3 antibody
- SIGLEC3 antibody
see all

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.
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.
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).
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).
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.
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)

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)

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)

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.

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab269456? Please let us know so that we can cite the reference in this datasheet.

ab269456 has not yet been referenced specifically in any publications.

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1-3 of 3 Abreviews or Q&A

Application

Immunocytochemistry

Sample

Human Cell (PBMCs)

Permeabilization

Specification

PBMCs

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: RT°C

Fixative

Acetone

Abcam user community

Verified customer

Submitted May 26 2021

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (PBMCs)

Permeabilization

Specification

PBMCs

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Fixative

Acetone

Abcam user community

Verified customer

Submitted Apr 22 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Spleen)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: EDTA

Permeabilization

Specification

Spleen

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 21°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Apr 08 2021

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As

Immunocytochemistry abreview for Anti-CD33 antibody [EPR23051-101]

Immunocytochemistry/ Immunofluorescence abreview for Anti-CD33 antibody [EPR23051-101]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD33 antibody [EPR23051-101]

Post-translational
modifications