Mouse Anti-CD28 Recombinant Antibody (clone ANC28.1/5D10); F(ab')2 Fragment (CAT#: DrMAB-981)

Recombinant Mouse Antibody binds selectively to Mouse CD28, expressed in HPB-M.


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Agonist

Figure 1 The CD28 superagonist ANC28.1 induces polyclonal T cell activation in vitro and leads to ZAP70 independent phosphorylation of LAT and PLCc1.

Figure 1 The CD28 superagonist ANC28.1 induces polyclonal T cell activation in vitro and leads to ZAP70 independent phosphorylation of LAT and PLCc1.

5x10⁴ freshly prepared human T cells per well were seeded in 96-well plates and treated with the indicated combinations of antibodies or were incubated in medium as a control.

Waibler, Z., Sender, L. Y., Merten, C., Hartig, R., Kliche, S., Gunzer, M., ... & Schraven, B. (2008). Signaling signatures and functional properties of anti-human CD28 superagonistic antibodies. PloS one, 3(3).

Stim

Figure 2 Induction of sustained calcium flux by human CD28 superagonistic antibody ANC28.1.

Figure 2 Induction of sustained calcium flux by human CD28 superagonistic antibody ANC28.1.

1x10⁶ freshly prepared Indo-1 loaded human T cells were stimulated with either a 1:50 v/v dilution of ascites fluid of CD3 mAb MEM92 (orange triangle), 10 mg/ml conventional CD28 mAb (CD28.2; blue triangle) followed by crosslinking with 20 mg/ml polyclonal goat anti-mouse antiserum (black triangle) or 10 mg/ml ANC28.1 (red triangle) followed by crosslinking with 20 mg/ml polyclonal goat anti-mouse antiserum (black triangle).

Waibler, Z., Sender, L. Y., Merten, C., Hartig, R., Kliche, S., Gunzer, M., ... & Schraven, B. (2008). Signaling signatures and functional properties of anti-human CD28 superagonistic antibodies. PloS one, 3(3).

Stim

Figure 3 Sustained calcium flux, T cell proliferation, and T cell signaling upon TGN1412 treatment.

Figure 3 Sustained calcium flux, T cell proliferation, and T cell signaling upon TGN1412 treatment.

5x10⁴ freshly prepared human T-cells/96-well were stimulated with the indicated concentrations of ANC28.1 or TGN1412 or were left untreated as a control. For crosslinking of ANC28.1, a polyclonal goat anti-mouse antiserum was used and crosslinking of TGN1412 was performed.

Waibler, Z., Sender, L. Y., Merten, C., Hartig, R., Kliche, S., Gunzer, M., ... & Schraven, B. (2008). Signaling signatures and functional properties of anti-human CD28 superagonistic antibodies. PloS one, 3(3).

Neu

Figure 4 Only engagement of CD81 by HCV-E2 or neutralizing Abs synergizes with ligation of CD28.

Figure 4 Only engagement of CD81 by HCV-E2 or neutralizing Abs synergizes with ligation of CD28.

Human naive T cells were cultured with recombinant HCV envelope proteins immobilized on plate-bound anti-E2 plus soluble anti-CD28 (clone ANC28.1/5D10).

Serra, A., Nuti, S., Tavarini, S., Sammicheli, C., Rosa, D., Saletti, G., ... & Wack, A. (2008). Coligation of the hepatitis C virus receptor CD81 with CD28 primes naive T lymphocytes to acquire type 2 effector function. The Journal of Immunology, 181(1), 174-185.

FC

Figure 5 ANC28 has properties of 'mitogenic' CD28 antibody.

Figure 5 ANC28 has properties of 'mitogenic' CD28 antibody.

CFSE-labeled PBMC stimulated with CD3, PHA, ANC28 or 28.2 alone, or CD28 mAb in combination with CD3 were analyzed for activation (CD25 up-regulation) and expansion (CFSE dye dilution) by flow cytometry on day 6.

Singh, M., Basu, S., Camell, C., Couturier, J., Nudelman, R. J., Medina, M. A., ... & Lewis, D. E. (2008). Selective expansion of memory CD4+ T cells by mitogenic human CD28 generates inflammatory cytokines and regulatory T cells. European journal of immunology, 38(6), 1522-1532.

FC

Figure 6 Selective expansion of CD4+CD45RO+ cells by ANC28.

Figure 6 Selective expansion of CD4+CD45RO+ cells by ANC28.

CFSE-labeled purified CD4⁺, CD8⁺ cells stimulated with 28.2 or ANC28.

Singh, M., Basu, S., Camell, C., Couturier, J., Nudelman, R. J., Medina, M. A., ... & Lewis, D. E. (2008). Selective expansion of memory CD4+ T cells by mitogenic human CD28 generates inflammatory cytokines and regulatory T cells. European journal of immunology, 38(6), 1522-1532.


Specifications

  • Immunogen
  • Recombinant Human CD28 and HPB-MLT Human tumor cells
  • Host Species
  • Mouse
  • Type
  • Mouse IgG1 F(ab')2, κ
  • Specificity
  • Antibody recognizes CD28 expressed on T cells and also on BHK cells transfected with Human CD28. This antibody has been characterized as a superagonist.
  • Species Reactivity
  • Human
  • Clone
  • ANC28.1/5D10
  • Applications
  • Agonist, Stim, Neu, FC

Product Property

  • Purification
  • Protein A purification
  • Purity
  • >95% as determined by analysis by SDS-PAGE
  • Concentration
  • 1 mg/ml
  • Buffer
  • 50 mM Sodium Phosphate pH 7.5, 100 mM Potassium Chloride, 150mM NaCl. Product was sterile filtered and vialed under aseptic conditions. No preservative added
  • 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
  • The CD28 antibody has been reported in applications of Agonist, Stimulation, Neutralization, Flow Cytometry.
    Agonist: 10 μg/ml.
    Stim: 10 mg/ml.

Target

  • Alternative Names
  • CD28; CD28 molecule; CD28 antigen (Tp44); T-cell-specific surface glycoprotein CD28; T cell specific surface glycoprotein; CD28 antigen; Tp44; MGC138290

Related Resources

  • Related Diseases

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-0424ZL Mouse Anti-CD28 Recombinant Antibody (clone CBLS-209) FC, IP, WB, IHC, FuncS Mouse IgG1
PABL-365 Mouse Anti-CD28 Recombinant Antibody (clone 1C6) WB, ELISA, FuncS Mouse IgG1
HPAB-0132CQ Human Anti-CD28 Recombinant Antibody (clone FR255734) ELISA, IHC, Block, FuncS Human IgG2M3, κ
HPAB-0245-FY Mouse Anti-CD28 Recombinant Antibody (HPAB-0245-FY) WB, IP, IF, FC Mouse IgG1
HPAB-0246-FY Mouse Anti-CD28 Recombinant Antibody (HPAB-0246-FY) FC, FuncS, ICC, IHC, IP, WB Mouse IgG1

Chimeric Antibody

CAT Product Name Application Type
TAB-068LC Anti-Human CD28 Recombinant Antibody (ChTN228) ELISA, FC Chimeric antibody (mouse/human)
TAB-068LC-S(P) Anti-Human CD28 Recombinant Antibody scFv Fragment (ChTN228) FC, ELISA Chimeric antibody (mouse/human)
TAB-068LC-F(E) Anti-Human CD28 Recombinant Antibody Fab Fragment (ChTN228) FC, ELISA Chimeric antibody (mouse/human)

Mouse Antibody

CAT Product Name Application Type
PABX-031 Recombinant Mouse Anti-CD28 Antibody (5.11A1) WB, ELISA, FuncS, Activ IgG
PABX-031-F (E) Recombinant Mouse Anti-CD28 Antibody Fab Fragment (5.11A1) WB, ELISA, FuncS Fab

Human Antibody

CAT Product Name Application Type
PABX-031-S (P) Recombinant Mouse Anti-CD28 Antibody scFv Fragment (5.11A1) WB, ELISA, FuncS scFv

Neutralizing Antibody

Blocking Antibody

CAT Product Name Application Type
NEUT-335CQ Hamster Anti-CD28 Recombinant Antibody (clone 37.51) IHC-Fr, FC, IP, Block, FuncS Hamster IgG
NEUT-336CQ Mouse Anti-CD28 Recombinant Antibody (clone E18) FC, Costim, Block Mouse IgG2b, κ

Rabbit Monoclonal Antibody

ADCC Enhanced Antibody

CAT Product Name Application Type
AFC-TAB-H47 Afuco™ Anti-CD28 ADCC Recombinant Antibody (Lumretuzumab), ADCC Enhanced ELISA, FC, IP, FuncS, IF, Neut ADCC enhanced antibody

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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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