Mouse Anti-ACKR1 Recombinant Antibody (clone NaM185-2C3 (2C3)) (CAT#: FAMAB-0008CQ)
Recombinant Mouse Antibody clone NaM185-2C3 (2C3), which is specific to ACKR1. The minimal DARC epitope recognized by 2C3 MAb is linear and consists of five amino acids: FEDVW.
Specific Inquiry
WB
Figure 1 Western blotting analysis of membranes proteins from normal and 4.1 (-) HE erythrocytes.
Membrane proteins from whole ghost lysates separated by SDS-PAGE were transferred to a nitrocellulose membrane and incubated with relevant primary antibodies against the indicated proteins. The following antibodies were used: anti 4.1R (PAb); anti-GPC (MAb 1F6); anti-Kell (MAb 5A11); anti XK (rabbit PAb); anti DARC (MAb NaM185-2C3); anti AQP1 (PAb AB3065); anti Rh (PAb MPC8); anti UT-B (rabbit PAb); anti RhAG (MAb LA18.18); anti band 3 (rabbit PAb); anti GAPDH (MAb). GADPH was used as loading control to show that similar amounts of membrane proteins were present on each lane. All experiments were repeated at least three times. HE, hereditary elliptocytosis; MAb, monoclonal antibody;PAb, polyclonal antibody
Azouzi, S., Collec, E., Mohandas, N., An, X., Colin, Y., & Le Van Kim, C. (2015). The human Kell blood group binds the erythroid 4.1 R protein: new insights into the 4.1 R‐dependent red cell membrane complex. British journal of haematology, 171(5), 862-871.
ELISA
Figure 2 Inhibition of 2C3 MAb binding to ECD1-nuc.
The antibody was incubated with serially diluted DFEDVWNSSYG (circles) or DFEDVWNSSY(SO3H)G (squares) peptides. Subsequent binding to ECD1-nuc was evaluated by ELISA. The difference in % of inhibition was statistically significant in the range of concentrations 3-60 nM (P < 0.05, two-tailed Mann-Whitney U test).
Smolarek, D., Hattab, C., Buczkowska, A., Kaczmarek, R., Jarząb, A., Cochet, S., ... & Grodecka, M. (2015). Studies of a murine monoclonal antibody directed against DARC: reappraisal of its specificity. PloS one, 10(2), e0116472.
ELISA
Figure 3 Comparison of binding of 2C3 MAb and 2C3 Fab to ECD1-nuc evaluated by ELISA.
The binding of 2C3 MAb (squares) and 2C3 Fab (circles) was evaluated by ELISA. The difference in binding is statistically significant at concentration 20 nM (P < 0.05, two-tailed Mann-Whitney U test).
Smolarek, D., Hattab, C., Buczkowska, A., Kaczmarek, R., Jarząb, A., Cochet, S., ... & Grodecka, M. (2015). Studies of a murine monoclonal antibody directed against DARC: reappraisal of its specificity. PloS one, 10(2), e0116472.
Specifications
- Immunogen
- CHO cells expressing the recombinant Duffy glycoprotein
- Host Species
- Mouse
- Type
- Mouse IgG1, κ
- Specificity
- Human ACKR1
- Species Reactivity
- Human
- Clone
- NaM185-2C3 (2C3)
- Applications
- ELISA, WB, FC, Block
Product Property
- Purity
- >95% as determined by SDS-PAGE and HPLC analysis
- Concentration
- Please refer to the vial label for the specific concentration.
- Buffer
- PBS
- Preservative
- No preservatives
- Storage
- Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.
Applications
- Application Notes
- Microtiter 96-well plate was coated with wild-type ECD1-nuc 0.1 μg of protein per well in 50 μl of carbonate buffer pH 9.6, overnight at 4°C. Nonspecific binding was blocked by incubation for 1 hour at room temperature with TBS (Trisbuffered saline: 20 mM Tris-HCl, and 150 mM NaCl pH 7.4) containing 5% non-fat dry milk. After washing the wells once with TBS, 50 μl of 2C3 MAb or 2C3 Fab fragments appropriately diluted in TBS were added to each well, and the mixture was incubated for 1 h at room temperature. After five washes with TBS, 50 μl of a 1000 x dilution in TBS of rabbit anti-Fab antiserum conjugated with alkaline phosphatase or avidin-alkaline phosphatase 1000 x diluted (Pierce, Rockford, IL, USA) were added to each well and the plates were incubated for 1 h at room temperature. To develop the reaction, p-nitrophenylphosphate was used (1 mg/ml in 60 mM carbonate buffer, pH 9.5, containing 1 mM MgCl₂); the OD₄₀₅ was determined using Perkin Elmer EnSpire 2300 Multilabel Reader. All experiments were performed in triplicates. Inhibition of the antibody binding was evaluated by incubating appropriately diluted MAb or Fab with serially diluted ECD1-nuc mutants for 1h. The samples were then applied on the coated plate and incubated for 10 min, and the bound antibody was detected. The percentage of inhibition was calculated as: [1- (OD₄₀₅ of sample)/(OD₄₀₅ of control)] x100.
Target
- Alternative Names
- Atypical Chemokine Receptor 1 (Duffy Blood Group); Duffy Blood Group, Atypical Chemokine Receptor; Duffy Blood Group, Chemokine Receptor; Plasmodium Vivax Receptor; Fy Glycoprotein; Glycoprotein D; DARC; GpFy; GPD; FY; Duffy Blood Group Chemokine Receptor; Duffy Antigen/Chemokine Receptor; Atypical Chemokine Receptor 1; Duffy Blood Group Antigen; Duffy Blood Group; CD234 Antigen; CCBP1; CD234; WBCQ1; Dfy
- Gene ID
- 2532
- UniProt ID
- Q16570
Product Notes
This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:
• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production
See more details about Hi-Affi™ recombinant antibody benefits.
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Recombinant Antibody
| CAT | Product Name | Application | Type |
|---|---|---|---|
| MOB-1354z | Mouse Anti-ACKR1 Recombinant Antibody (clone 9C5) | WB, ELISA, FC, ICC, IF | Mouse IgG1, κ |
| FAMAB-0007CQ | Human Anti-ACKR1 Recombinant Antibody (clone ch2C3) | ELISA, FC, Block | Chimeric (mouse/human) IgG4, κ |
| FAMAB-1017CQ | Human Anti-ACKR1 Recombinant Antibody (clone CA52) | Block, SPR, WB, ELISA | Human IgG1 |
| FAMAB-1352CQ | Mouse Anti-ACKR1 Recombinant Antibody (clone Hima-19) | ELISA | Mouse IgG1 |
Single-domain Antibody
| CAT | Product Name | Application | Type |
|---|---|---|---|
| NABG-008 | Recombinant Anti-Human ACKR1 VHH Single Domain Antibody | ELISA, IHC, FC, FuncS | Llama VHH |
Rabbit Monoclonal Antibody
| CAT | Product Name | Application | Type |
|---|---|---|---|
| MOR-0927 | Hi-Affi™ Rabbit Anti-ACKR1 Recombinant Antibody (clone DS927AB) | WB, IHC, ICC, IP, FC | Rabbit IgG |
Fab Fragment Antibody
| CAT | Product Name | Application | Type |
|---|---|---|---|
| FAMAB-0007CQ-F(E) | Human Anti-ACKR1 Recombinant Antibody (clone ch2C3); Fab Fragment | ELISA, FC, Block | Chimeric (mouse/human) Fab |
| FAMAB-0008CQ-F(E) | Mouse Anti-ACKR1 Recombinant Antibody (clone NaM185-2C3 (2C3)); Fab Fragment | ELISA, FC, Block | Mouse Fab |
scFv Fragment Antibody
| CAT | Product Name | Application | Type |
|---|---|---|---|
| FAMAB-0008CQ-S(P) | Mouse Anti-ACKR1 Recombinant Antibody (clone NaM185-2C3 (2C3)); scFv Fragment | ELISA, FC, Block | Mouse scFv |
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For Research Use Only. Not For Clinical Use.
For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.
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