Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22238] to BAFF
- Suitable for: Flow Cyt, ICC/IF, IP, WB
- Reacts with: Human
Product nameAnti-BAFF antibody [EPR22238]
See all BAFF primary antibodies
DescriptionRabbit monoclonal [EPR22238] to BAFF
Tested applicationsSuitable for: Flow Cyt, ICC/IF, IP, WBmore details
Unsuitable for: IHC-P
Species reactivityReacts with: Human
Recombinant fragment within Human BAFF aa 50 to the C-terminus. The exact sequence is proprietary.
Database link: Q9Y275
- WB: U937 and HL-60 whole cell lysates. ICC/IF: U937 cells. Flow Cyt: HL-60 cells. IP: U937 whole cell lysate.
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
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.
Storage instructionsShipped 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 bufferpH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab224710 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Detects a band of approximately 30, 32 kDa (predicted molecular weight: 31 kDa).|
FunctionCytokine that binds to TNFRSF13B/TACI and TNFRSF17/BCMA. TNFSF13/APRIL binds to the same 2 receptors. Together, they form a 2 ligands -2 receptors pathway involved in the stimulation of B-and T-cell function and the regulation of humoral immunity. A third B-cell specific BAFF-receptor (BAFFR/BR3) promotes the survival of mature B-cells and the B-cell response.
Tissue specificityAbundantly expressed in peripheral blood Leukocytes and is specifically expressed in monocytes and macrophages. Also found in the spleen, lymph node, bone marrow, T-cells and dendritic cells. A lower expression seen in placenta, heart, lung, fetal liver, thymus, and pancreas.
Sequence similaritiesBelongs to the tumor necrosis factor family.
modificationsThe soluble form derives from the membrane form by proteolytic processing.
Cellular localizationSecreted and Cell membrane.
- Information by UniProt
- ApoL related ligand TALL 1 antibody
- B cell Activating Factor antibody
- B lymphocyte stimulator antibody
All lanes : Anti-BAFF antibody [EPR22238] (ab224710) at 1/1000 dilution
Lane 1 : U937 (human histiocytic lymphoma cell line) whole cell lysate
Lane 2 : HL-60 (human promyelocytic leukemia cell line) whole cell lysate
Lane 3 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate
Lane 4 : Daudi (human Burkitt's lymphoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 31 kDa
Observed band size: 30,32 kDa why is the actual band size different from the predicted?
Exposure time: 26 seconds
Blocking/Dilution buffer: 5% NFDM/TBST.
BAFF was immunoprecipitated from 0.35 mg of U937 (human histiocytic lymphoma cell line) whole cell lysate with ab224710 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224710 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: U937 whole cell lysate 10 µg (Input).
Lane 2: ab224710 IP in U937 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab224710 in U937 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 20 seconds.
Flow cytometric analysis of Daudi (human Burkitt's lymphoma cell line; left panel) and HL-60 (human promyelocytic leukemia cell line ;right panel) cell lines labeling BAFF with ab224710 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Negative control: Daudi. (PMID: 20951740)
Gated on viable cells.
Immunofluorescent analysis of 100% methanol-fixed U937 (human histiocytic lymphoma cell line) cells labeling BAFF with ab224710 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on U937 cells.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
Confocal image showing membranous staining in U-937 cells.
We don’t recommend PFA fixation, since the result shows some background staining in the nuclear.
Negative control: HEK-293（PMID: 17393395.
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.
ab224710 has not yet been referenced specifically in any publications.