Mouse Anti-KLH IgM(Keyhole Limpet Hemocyanin IgM) ELISA Kit

This kit is based on capture ELISA detection method and takes 3h assay time. The microplate provided in this kit has been precoated with anti Mouse IgM(U-chain). HRP-antigen is used as detection antigen. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add HRP-antigen. Then, it binds with Anti-KLH IgM bound to anti Mouse IgM(U-chain). Wash unbound components and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of Anti-KLH IgM in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

Catalogue No.: 
Anti-KLH IgM ELISA Kit, Keyhole Limpet Hemocyanin IgM ELISA Kit
Product Name
Mouse Anti-KLH IgM(Keyhole Limpet Hemocyanin IgM) ELISA Kit
Anti-KLH IgM ELISA Kit, Keyhole Limpet Hemocyanin IgM ELISA Kit
Catalogue No.
UniProt No.
CAS: 9013-72-3
Sample Type
Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
Detection Method
Capture ELISA
Detection Wavelength
Reaction Duration
3 hours
2-8°C(Sealed), Don't cryopreserve.
Specifically binds with Anti-KLH IgM , no obvious cross reaction with other analogues.
ELISA Kit Components
Kit Components Item Size(48T) Size(96T) Storage Condition for Opened Kit
E001 ELISA Microplate(Dismountable) 8×6 8×12 Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E002 Lyophilized Standard 1vial 2vial Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E003 HRP-Antigen(Concentrated, 100X) 60ul 120ul 2-8°C (Avoid Direct Light)
E024 TMB Substrate 5ml 10ml
E039 Sample Dilution Buffer 10ml 20ml 2-8°C
E040 Antigen Dilution Buffer 5ml 10ml
E026 Stop Solution 5ml 10ml
E038 Wash Buffer(Concentrated, 25X) 15ml 30ml
E006 Plate Sealer 3 pieces 5 pieces  
E007 Product Description 1 copy 1 copy
Required Instruments and Reagents
  1. Microplate reader (wavelength: 450nm)
  2. 37°C incubator (CO2 incubator for cell culture is not recommenced.)
  3. Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
  4. Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
  5. Sterile tubes and Eppendorf tubes with disposable tips
  6. Absorbent paper and loading slot
  7. Deionized or distilled water
Standard Curve

This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)

Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(ng/ml) OD-1 OD-2 Average
0 0.078 0.082 0.08
1.562 0.144 0.151 0.147
3.125 0.204 0.214 0.208
6.25 0.297 0.312 0.303
12.5 0.48 0.505 0.49
25 0.786 0.826 0.802
50 1.44 1.513 1.469
100 2.173 2.284 2.217
EM2511-CM Standard Curve Image

Add a certain amount of Anti-KLH IgM into the sample. Calculate the recovery by comparing the measured value with the expected amount of Anti-KLH IgM in the sample.

Sample Type Recovery Range(%) Average(%)
serum(n=10) 89-104 95
EDTA plasma(n=10) 90-101 95
Heparin plasma(n=10) 89-101 96

Dilute the sample with a certain amount of Anti-KLH IgM at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type 1:2 1:4 1:8
serum(n=10) 89-105% 85-97% 90-103%
EDTA plasma(n=10) 89-99% 84-101% 82-97%
Heparin plasma(n=10) 84-96% 81-98% 82-91%

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.

Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/ml) 3.07 12.97 48.53 3.09 12.55 49.84
Standard deviation 0.19 0.7 2.44 0.17 0.63 2.53
CV(%) 6.2 5.43 5.02 5.43 5.02 5.08

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5) 37°C for 1 month 2-8°C for 6 months
Average(%) 80 95-100