Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µl 298 185 ₸

Order now and get it on Tuesday March 02, 2021

Anti-CD22 antibody [EPR20061] (ab207727)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20061] to CD22
Suitable for: WB, IHC-P, ICC/IF, IP
Reacts with: Human

Product name

Anti-CD22 antibody [EPR20061]
See all CD22 primary antibodies
Description

Rabbit monoclonal [EPR20061] to CD22
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Ramos, Daudi and Raji whole cell lysates; Human tonsil and fetal spleen lysates. IHC-P: Human spleen and tonsil tissues. ICC/IF: Daudi and Raji cells. IP: Human tonsil lysate; Raji whole cell lysate.
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20061
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD22 antibody [EPR20061] (ab207727)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling CD22 with ab207727 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membranous and cytoplasmic staining on B cells of human spleen [PMID: 11967115]. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-CD22 antibody [EPR20061] (ab207727)

Immunofluorescent analysis of 100% methanol-fixed Daudi (Human Burkitt's lymphoma cell line) cells labeling CD22 with ab207727 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Daudi cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Western blot - Anti-CD22 antibody [EPR20061] (ab207727)

Lanes 1 & 4 : Anti-CD22 antibody [EPR20061] (ab207727) at 1/2000 dilution
Lanes 2-3 : Anti-CD22 antibody [EPR20061] (ab207727) at 1/10000 dilution

Lane 1 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 3 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 4 : Human tonsil 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: 95 kDa
Observed band size: 130 kDa
why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure times: Lane 1: 3 seconds; Lane 2/3: 1 second; Lane 4: 8 seconds.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 24264377 & 21178016).
Immunoprecipitation - Anti-CD22 antibody [EPR20061] (ab207727)

CD22 was immunoprecipitated from 0.35 mg of Human tonsil lysate with ab207727 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207727 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: Human tonsil lysate, 10 μg (Input).

Lane 2: ab207727 IP in Human tonsil lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207727 in Human tonsil lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD22 antibody [EPR20061] (ab207727)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD22 with ab207727 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membranous and cytoplasmic staining on B cells of human tonsil [PMID: 11967115]. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-CD22 antibody [EPR20061] (ab207727)

Immunofluorescent analysis of 100% methanol-fixed Raji (Human Burkitt's lymphoma cell line) cells labeling CD22 with ab207727 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Raji cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Western blot - Anti-CD22 antibody [EPR20061] (ab207727)

Anti-CD22 antibody [EPR20061] (ab207727) at 1/1000 dilution + Human fetal spleen lysate at 10 µg

Secondary
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

Predicted band size: 95 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 24264377 & 21178016).
Immunoprecipitation - Anti-CD22 antibody [EPR20061] (ab207727)

CD22 was immunoprecipitated from 0.35 mg of Raji (Human Burkitt's lymphoma cell line) whole cell lysate with ab207727 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207727 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: Raji whole cell lysate, 10 μg (Input).

Lane 2: ab207727 IP in Raji whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207727 in Raji whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD22 antibody [EPR20061] (ab207727)

Immunohistochemical analysis of paraffin-embedded human thymoma tissue labeling CD22 with ab207727 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative control: no staining on human thymoma [PMID: 11967115].

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-CD22 antibody [EPR20061] (ab207727)

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