Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸ 10 µl 110 563 ₸

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Anti-CD19 antibody [EPR23174-145] (ab245235)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23174-145] to CD19
Suitable for: WB, Flow Cyt, IP, IHC-Fr, IHC-P, ICC/IF
Reacts with: Mouse

Product name

Anti-CD19 antibody [EPR23174-145]
See all CD19 primary antibodies
Description

Rabbit monoclonal [EPR23174-145] to CD19
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse
IHC-P	Mouse
IP	Mouse
WB	Mouse

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Mouse spleen tissue lysate. WEHI-231 and A20 whole cell lysate. IHC-P: Mouse spleen tissue. ICC/IF: Mouse peripheral blood mononuclear cells. Flow Cyt: Mouse peripheral blood mononuclear cells. IP: Mouse spleen lysate.IHC-Fr: Mouse spleen tiissue.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

EPR23174-145
Isotype

IgG
Research areas

Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized perfusion fixed frozen Mouse spleen tissue labeling CD19 with ab245235 at 1/250 (1.82 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse spleen. is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)

All lanes : Anti-CD19 antibody [EPR23174-145] (ab245235) at 1/1000 dilution

Lane 1 : WEHI-231 (mouse B cell lymphoma B lymphocyte), whole cell lysate
Lane 2 : A20 (mouse reticulum sarcoma B lymphocyte), whole cell lysate
Lane 3 : CTLL-2 (mouse 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: 60 kDa
Observed band size: 60-120 kDa
why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBS.

Exposure time: 48 seconds.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 26516065).

Negative control: CTLL-2 (PMID: 1370948) is treated as a negative cell line according to the literature, that CD19 is a B lymphocyte cell-surface marker that is expressed early during pre-B-cell differentiation, not in T cells.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [EPR23174-145] (ab245235)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD19 with ab245235 at 1/1000 dilution (0.446 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Positive staining on mouse spleen (PMID: 18648532). The section was incubated with ab245235 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunocytochemistry - Anti-CD19 antibody [EPR23174-145] (ab245235)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse PBMC (mouse primary peripheral blood mononuclear cell) cells labeling CD19 with ab245235 at 1/50 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in some subsets of mouse PBMCs. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Flow Cytometry - Anti-CD19 antibody [EPR23174-145] (ab245235)

Flow cytometric analysis of mouse peripheral blood mononuclear cell (PBMC) cells labeling CD19 with ab245235 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Cells were stained with rabbit IgG (Left) or ab245235 (Right). Then stained with anti-CD3 conjugated to Alexa Fluor^® 647. CD19 and CD3 are mutually exclusive expressed in mouse PBMC.

Gated on viable cells.
Immunoprecipitation - Anti-CD19 antibody [EPR23174-145] (ab245235)

CD19 was immunoprecipitated from 0.35 mg mouse spleen lysate with ab245235 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab245235 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: Mouse spleen lysate 10µg

Lane 2: ab245235 IP in mouse spleen lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245235 in mouse spleen lysate

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

Exposure time: 30 seconds.
Western blot - Anti-CD19 antibody [EPR23174-145] (ab245235)

Anti-CD19 antibody [EPR23174-145] (ab245235) at 1/1000 dilution + Mouse spleen tissue lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 60 kDa
Observed band size: 60-120 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 48 seconds.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 26516065).
Anti-CD19 antibody [EPR23174-145] (ab245235)

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