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

• ELISA

Supplier Data

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.

• FuncS

Lab

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

Unknown

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

Unknown

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

Unknown

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).

• ELISA

Unknown

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.

• WB

Lab

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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 280 kDa

false

Exposure time: 120s

• WB

Supplier Data

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

false