Anti-Polyethylene glycol antibody [PEG-B-47]

8 product images

• 

  ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

  ELISA Graph generated using purified ab51257 at 1μg/mL.
  

  Antigen concentration range:

  PEG5000 and PEG20000: 0.0078 - 0.5 μg/mL
  ADI-PEG: 0.0156 - 1 μg/mL
  PEG-BSA: 0.0039 - 0.25μg/mL
  PEG-IFN-α-2b0.0156 - 1 μg/mL

  Secondary antibody was an Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG(H+L) at 1/2500.

• 

  Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

  Blocking/Diluting buffer and concentration 5% NFDM/TBST

  Lane 1: Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257) at 1/1000 dilution

  Lane 2: Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257) at 1/5000 dilution

  All lanes: Bovine serum albumin (BSA) and PEG 5000 conjugated protein at 0.015 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 280 kDa

  Exposure time: 120s

• 

  ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

  Comparison of sandwich ELISA using RabMAb/RabMAb (ab51257/ab51257) and MAb/RabMAb (Vendor A/ab51257) for capture/detection.
  

  ab51257/ab51257*: Plate coated with 5 ug/mL of #47; 5 ug/mL of #47 used for detection (*Anti-PEG 47 biotin labeled).

  Vendor A/ ab51257*: Plates coated with 100 ug/mL of Vendor A Mouse MAb; 5 ug/mL of #47 used for detection (*Anti-PEG 47 biotin labeled).

• 

  Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

  Blocking buffer and concentration: 5% NFDM/TBST.

  Diluting buffer and concentration: 5% NFDM /TBST.

  All lanes: Western blot - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257) at 0.015 µg/mL

  All lanes: BSA-PEG at 10 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution

• 

  Functional Studies - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

  Method：Protein A sensor + Antigen (PEG 5K-BSA at 0.3μg/ml)+ antibody (ab51257 at 0.0312, 0.0625, 0.125, 0.25, 0.5, 1 μg/m).
  

  Results:

  Sample ID: PEG-5K-BSA

  KD (M): 2.41E-09

  kon(1/Ms): 2.67E+04

  kon Error: 6.81E+02

  kdis(1/s): 6.43E-05

  kdis Error: 2.92E-06

  Full X^2: 0.018444

  Full R^2: 0.982059

• 

  ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

  Comparison of 10 ug/well of activated linear (PEG5K) and branched (PEG40K) PEG using 5 ug/ml of ab51257 in Direct ELISA assay.
  

  Accuracy: By detecting the methoxy group of the PEG molecule itself, ab51257 is useful in measuring the pharmacokinetics of PEG-modified molecules in vivo. Data indicate that ab51257 detects various length Y-chain PEG molecules as well as single chain PEG molecules with equal affinity. ab51257 does not cross react with non-specific targets in blood or serum.

• 

  ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

  ELISA assay using ab51257 to detect different forms of PEG. PEG40000-BSA is a 40 kDa PEG molecule attached to BSA. PEG5000-BSA is a 5 kDa linear PEG molecule attached to BSA.

• 

  ELISA - Anti-Polyethylene glycol antibody [PEG-B-47] (ab51257)

  Comparison of ab51257 and Vendor A mouse MAb in Direct ELISA assy.
  
  Goat anti-rabbit IgG-AP used for anti-PEG-47 detection; goat anti-mouse IgM-AP used for Vendor A MAb detection.

  
  
  Fig 1a. Direct ELISA using 1 ug/mL of PEG-GCSF.

  
  
  Fig 1b. Direct ELISA using 1 ug/mL of PEG-IFN.

  Comparison to other anti-PEG: In both direct and sandwich ELISA assays, ab51257 shows greater affinity and accuracy than other anti-PEG antibodies when determining the concentration of PEG or PEG-modified proteins. Results were similar whether detecting PEG itself or PEG-modified targets.