Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 02, 2021

Anti-CD16 antibody [EPR22409-124] (ab246222)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22409-124] to CD16
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
Reacts with: Human

Product name

Anti-CD16 antibody [EPR22409-124]
See all CD16 primary antibodies
Description

Rabbit monoclonal [EPR22409-124] to CD16
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
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: Human spleen and colon cancer lysates. IP: Human spleen lysate. IHC-P: Human spleen and liver tissue. ICC/IF: Human PBMCs. Flow Cyt: Human PBMCs.
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

EPR22409-124
Isotype

IgG
Research areas

Immunoprecipitation - Anti-CD16 antibody [EPR22409-124] (ab246222)

CD16 was immunoprecipitated from 0.35 mg of human spleen lysate with ab246222 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab246222 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as the secondary antibody at 1/5000 dilution.

Lane 1: Human spleen lysate 10 µg (Input).
Lane 2: ab246222 IP in human spleen lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab246222 in human spleen lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
Flow Cytometry - Anti-CD16 antibody [EPR22409-124] (ab246222)

Human PBMCs (human peripheral blood mononuclear cells) were stained with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left) or ab246222 (Right) followed by a Goat anti rabbit IgG (Dylight ^® 488) at 1/2000 dilution. They were then stained with Alexa Fluor^® 647-conjugated anti-CD14. The expression pattern is consistent with that described in the literature. (PMID: 21738687)

Gated on viable cells.
Flow Cytometry - Anti-CD16 antibody [EPR22409-124] (ab246222)

Negative control: Raji. (PMID:11207281).

Raji (human Burkitt's lymphoma cell line) were stained with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)(Left) or ab246222 (Right) followed by a Goat anti rabbit IgG (Dylight ^® 488) at 1/2000 dilution.

Gated on viable cells.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 antibody [EPR22409-124] (ab246222)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD16 with ab246222 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Kuffer cells of human liver (PMID: 26512139) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 antibody [EPR22409-124] (ab246222)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling CD16 with ab246222 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on red pulp of human spleen (PMID: 26512139; 29692344). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-CD16 antibody [EPR22409-124] (ab246222)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized human PBMCs (human peripheral blood mononuclear cells) or Raji (human Burkitt's lymphoma cell line) cell line labeling CD16 with ab246222 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 membranous staining in subsets of human PBMC cells is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

Negative control: Raji (PMID:11207281).
Western blot - Anti-CD16 antibody [EPR22409-124] (ab246222)

All lanes : Anti-CD16 antibody [EPR22409-124] (ab246222) at 1/1000 dilution

Lane 1 : Human spleen lysate
Lane 2 : Human colon cancer lysate
Lane 3 : Raji (human Burkitt's lymphoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/100000 dilution

Predicted band size: 29 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times.

Lane 1: 92 seconds.

Lanes 2 & 3: 3 minutes.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 2526846).

Negative control: Raji (PMID:11207281).
Anti-CD16 antibody [EPR22409-124] (ab246222)

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