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Vial / BottleCapLabelFunction/DescriptionContent
1whiteResidual Protein Galactosyltransferase, Incubation Buffer
  • Ready-to-use solution
  • For preparation of the Biotin DIG working Solution
1 bottle,100 ml
2blueResidual Protein Galactosyltransferase, Conjugate Buffer
  • Ready-to-use solution
  • For preparation of the Anti-DIG POD Reagent, Vial 6
1 bottle,100 ml
3yellowResidual Protein Galactosyltransferase, BiotinConjugate
  • Biotin labeled for target capture
  • Monoclonal anti-1,4-Galactosyltransferase antibody
  • 20-fold stock solution
1 vial,0.75 ml
4purpleResidual Protein Galactosyltransferase, DIG conjugate
  • Digoxigenin labeled for target marking
  • Monoclonal anti-1,4-Galactosyltransferase antibody
  • 20-fold stock solution
1 vial,0.75 ml
5redResidual Protein Galactosyltransferase, Wash Buffer
  • For washing steps
  • 10-fold stock solution
1 bottle,100 ml
6redResidual Protein Galactosyltransferase, Anti-DIG PODReagent
  • Peroxidase conjugated to anti-DIG antibody
  • Lyophilizate
1 bottle
7blackResidual Protein Galactosyltransferase, Detection Substrate (TMB)
  • For color development nd detection
  • Ready-to-use solution
1 bottle,15 ml
8colorlessResidual Protein Galactosyltransferase, Stop Solution
  • For stopping the color development
  • Ready-to-use solution
1 bottle,15 ml
9light brownResidual Protein Galactosyltransferase, Standard A
  • For calibration of the assay
  • Lyophilizate
  • 1,4-Galactosyltransferase free
1 bottle
10light brownResidual Protein Galactosyltransferase, Standard B
  • For calibration of the assay
  • 1,4-Galactosyltransferase
  • Lyophilizate
1 bottle
11light brownResidual Protein Galactosyltransferase, Standard C
  • For calibration of the assay
  • 1,4-Galactosyltransferase
  • Lyophilizate
1 bottle
12light brownResidual Protein Galactosyltransferase, Standard D
  • For calibration of the assay
  • 1,4-Galactosyltransferase
  • Lyophilizate
1 bottle
13light brownResidual Protein Galactosyltransferase, Standard E
  • For calibration of the assay
  • 1,4-Galactosyltransferase
  • Lyophilizate
1 bottle
14light brownResidual Protein Galactosyltransferase, Standard F
  • For calibration of the assay
  • 1,4-Galactosyltransferase
  • Lyophilizate
1 bottle
15whiteResidual Protein Galactosyltransferase, Control X
  • Positive control
  • 1,4-Galactosyltransferase
  • Lyophilizate
1 bottle
16greenResidual Protein Galactosyltransferase, Control Y
  • Positive control
  • 1,4-Galactosyltransferase
  • Lyophilizate
1 bottle
17-Residual Protein Galactosyltransferase, Coated Micro Titer Plate
  • Target capture
  • Streptavidin- coated multi-well plate
  • Ready-to-use
12 strips in frame
18-Residual Protein Galactosyltransferase, Multi-well SealingFilm
  • Sealing MWP during
  • incubations
10 pieces of self-adhesive film
", "Name": "Content" }, { "Language": "en", "Value": "
  • Polypropylene tubes for preparation of antibody reagents
  • 1 liter bottle for wash buffer preparation
  • Micropipettes
  • Centrifuge
  • Multi-well plate shaker
  • Roller mixer
  • ELISA reader
  • Multi-well plate washer
", "Name": "Additional Equipment and Reagent Required" }, { "Language": "en", "Value": "
  • Incubation time: 1 hour 45 minutes.
  • Total assay time: approximately 2 hours 30 minutes.
", "Name": "Assay Time" }, { "Language": "en", "Value": "
  • Aqueous buffer solutions from biotechnology processes.
  • Cell culture supernatant
", "Name": "Sample Materials" }, { "Language": "en", "Value": "
Always ensure that positive and negative controls are tested on each plate together with samples.
Positive controls
Control X and Y are positive controls and are provided with the kit. For preparation of the positive controls, see section Preparation of Working Solutions.
Negative control
Analyte-free matrix of your sample material, such as buffer or cell culture medium.", "Name": "Control Reactions" }, { "Language": "en", "Value": "A quantitative, functional assay is performed to assess background signal, and the lot-specific concentrations of Standards A to F and Controls X and Y.", "Name": "Quality Control" }, { "Language": "en", "Value": "
ObservationPossible causeRecommendation
Absorbance of samples too low  \t\t\t\t\t  Insufficient TMB reactionIncrease the incubation time with the Detection Substrate (TMB) for color development (Step 12). The incubation time can be increased until the wells with the higher concentrated standards show a clearly observable blue color, up to 30 minutes.
POD conjugate has a reduced enzymatic activity.Use freshly prepared POD conjugate.
Low concentration after dilution of highly concentrated samples\t\t\t\t\t\t\t\t\t\t\t   Reduce dilution factor for highly concentrated samples.
Absorbance ofsamples too highConcentration exceeds measuring range.Dilute samples to ensure that they are within the specified detection range.
Background of sample matrix.Check analyte-free sample matrix for background absorbance value. Correct sample values for background absorbance of matrix.
Contamination of samples with 1,4-Galactosyltransferase from lab workspace.Check absorbance values of Standard A and negative control. If the absorbance values of Standard A and the negative control are significantly increased, repeat the experiment in a different workspace using a new kit. Be careful when reconstituting lyophilizates. Thoroughly clean contaminated workspace.
Absorbance ofnegative controltoo highContamination of matrix used as negative control with 1,4-Galactosyltransferase from lab workspace.Check for potential contamination with 1,4-Galactosyltransferase in the analyte-free matrix. Use a matrix sample from an independent batch or another 1,4-Galactosyltransferase-free solution (e.g. water) for comparison.
Absorbance of the background too highSubstrate shows color development without enzymatic activity.Use a newly opened vial of Detection Substrate (TMB).
Variations toohighInsufficient mixing of sample and incubation mix.Ensure mixing on the multi-well plate mixer is done with 300 rpm.
Residual buffer after washingCheck performance of multi-well plate washer. Wells should not contain residual buffer after washing. Try tapping the Multi-well Plate on an absorbent paper towel to remove traces of Wash Buffer.
Sample matrix difficult to pipette due to viscosity or composition.Carefully pipette samples without droplets on outside of tip. Avoid high aspiration or ejection velocity when pipetting the samples.
Precision of micropipettesCheck the precision of the micropipettes.
Multi-well plate washer not washing correctly.Check the multi-well plate washer for tip blockage or salt crystallization, which may affect the evenness and effectiveness of the washing steps.
", "Name": "Troubleshooting" }, { "Language": "en", "Value": "
Use double-distilled or deionized water of equivalent quality, for reconstitution of the lyophilizates. Ensure that the lyophilizates are carefully reconstituted to avoid contamination of the environment. Contamination of the workspace may potentially contaminate samples to be tested.
A roller mixer may be used to effectively dissolve lyophilizates
In addition to the ready-to-use solutions supplied with this kit, you will need to prepare the following working solutions:
SolutionContentReconstitution/Preparation of Working SolutionStorage and StabilityFor use in...
9 to 14Standards A to F (Bottles 9 to 14)Add 500 μl of double-distilled water to each of the vials and mix thoroughly for 30 minutes to completely dissolve the lyophilizate.Store 4 weeks at +2 to +8°CCalibration of the assay
15 and 16Controls X and Y (Bottles 15 and 16)
  • Add 500 μl of double-distilled water to each of the vials and mix thoroughly for 30 minutes to completely dissolve the lyophilizate.
  • The controls supplied with the kit are ready to use once reconstituted.
Do not further dilute the Positive Controls X and Y.
Store 4 weeks at +2 to +8°CPositive controls
5Wash Buffer (Bottle 5)Dilute the entire contents of the bottle (100 ml) with 900 ml water and mix thoroughly.Store 4 weeks at +2 to +8°CWashing steps
3Biotin Conjugate (Vial 3)
  • Dilute the appropriate volume 1:10 with Incubation Buffer from Bottle 1.
  • For a 96-well plate, prepare 500 μl of Biotin Conjugate stock solution + 4.5 ml of Incubation Buffer.
  • If using only a part of the 96 wells, then prepare a proportionally reduced volume of this working solution.
Store 4 weeks at +2 to +8°CPreparation of Biotin DIG Working SolutionTarget capture
4DIG Conjugate (Vial 4)
  • Dilute the appropriate volume 1:10 with Incubation Buffer from Bottle 1.
  • For a 96-well plate, prepare 500 μl of DIG Conjugate stock solution + 4.5 ml of Incubation Buffer.
  • If using only a part of the 96 wells, then prepare a proportionally reduced volume of this working solution.
Store 4 weeks at +2 to +8°CPreparation of Biotin DIG Working SolutionTarget marking
6Anti-DIG POD Reagent (Bottle 6)
  1. Add 500 μl double-distilled water to the vial and mix thoroughly for 30 minutes to completely dissolve the lyophilizate. The concentration of the reconstituted solution is 4 U/ml.
  2. Dilute the 4 U/ml solution with Conjugate Buffer (Bottle 2) to achieve a working concentration of 10 mU/ml, as follows:
  • 25 μl Anti-DIG POD Solution (4 U/ml) + 9.975 ml Conjugate Buffer.   
Store 4 weeks at +2 to +8°CDetection
-Biotin DIG Working SolutionCombine equal volumes of the prepared working solutions: Biotin Conjugate (Solution 3) and DIG Conjugate (Solution 4).Mix thoroughly, for example, using a roller mixer for 10 minutes.
Always prepare the Biotin DIG Working Solution just before use, and use the solution within 30 minutes after preparation.
Use within 30 minutes after preparation.ELISA Step 6
", "Name": "Working Solution" }, { "Language": "en", "Value": "The Roche Residual Protein Galactosyltransferase kit, based on an immuno assay with tailor-made monoclonal antibodies against β-1,4-Galactosyltransferase makes testing for residual β-1,4-Galactosyltransferase, rec. sensitive and reliable. The assay is suited for process development, in-process control and final quality control.", "Name": "Product Description" }, { "Language": "en", "Value": "Immunologic assay for sensitive detection of β-1,4-Galactosyltransferase.", "Name": "Positioning" }, { "Language": "en", "Value": "
  • Fast: Total assay time of 2 - 2.5 hours
  • High sensitivity: Detection limit ≤ 0.25 ng/mL, measuring range 0.25 - 25 ng/mL
  • High specificity: Specific detection of β-1,4-Galactosyltransferase by use of monoclonal antibodies
", "Name": "Benefits" }, { "Language": "en", "Value": "The kit is designed for sensitive detection of residual β-1,4-Galactosyltransferase in in-process control and release testing.", "Name": "Applications" }, { "Language": "en", "Value": "Performance: Corresponds to specification
Stability: At +2 to 8°C within specification for 12 months.", "Name": "Specification" } ] } } ] }

Residual Protein Galactosyltransferase

ELISA kit for quantitative detection of β-1,4-Galactosyltransferase

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