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Vitamin B12(VB12)

Antibody

Antibody

Name Vitamin B12 Antibody (VB12 Antibody)
Catalog No. H835e1
Clonality Monoclonal
Host Rabbit
Reactive Species Human
Application CLIA(AE) CLIA(ALP) ICA
C1770 (Capture) - H835e1 (Detection) H835e1 (Capture) - C1716 (Detection) H835e1 (Capture) - C1750 (Detection)
Purity Protein A/G Purified, Purity >95%
Storage Buffer 1xPBS,pH 7.4
Storage Instructions Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Antigen

Antigen

Name Intrinsic Factor (IF)
Catalog No. C1716 C1750 C1770
Species Swine
Type Recombinant
Tag His
Expression System Eukaryotic cell
SDS-PAGE C1716-SDS-PAGE C1750-SDS-PAGE C1770-SDS-PAGE
Purity >90% (R250-stained SDS-PAGE)
Storage Buffer 1xPBS,pH7.4
Storage Instructions Store at -80°C in aliquots. Avoid repeated freeze-thaw cycles.
Antigen

CLIA Bulk Reagent

Name Catalog No. Recommended Platform Methodology
Vitamin B12 (VB12) Bulk Reagent Q801a1 CLIA(AE) Sandwich Assay
Q801a2 CLIA(ALP)
VB12

Nanjing OkayBio launches its innovative Vitamin B12 (VB12) sandwich assay series, featuring a groundbreaking 3-component chemiluminescent immunoassay(CLIA) bulk reagent. This advancement streamlines traditional 5-component systems, reducing detection time by 30% through optimized reaction protocols. Validated on CLIA platforms, the reagent achieves a sensitivity of?5 pg/mL?and demonstrates exceptional correlation (R2 >0.95) with Roche-assigned samples.

VB12
VB12

Product Data

Product Data

1.Calibration Curve

VB12 antibody (H835e1) and intrinsic factor (C1770) were used to detect VB12 samples (0–2000 pg/mL).

VB12 Calibration Curve

Figure 1. VB12 Calibration Curve

Sample Concentration (pg/mL) RLU
1 0.0 8308
2 22.2 64128
3 52.3 109666
4 499.5 957793
5 1004.1 2084186
6 2015.4 4067850

Table 1. VB12 Calibration Curve Data

2.Sample Concordance

Roche-assigned samples (35.1–1934 pg/mL, n=48 ) tested with H835e1 and C1770 showed?R2 = 0.9926.

VB12 Sample Concordance 
(Roche-assigned)

Figure 2. VB12 Sample Concordance
(Roche-assigned)

Sample Assigned
Conc.(pg/mL)
Detected
Conc.(pg/mL)
1 35.1 43.8
2 36.2 32.4
3 54.3 59.8
? ? ?
47 1872 1931
48 1934 1889

Table 2. VB12 Sample Concordance Data
(Roche-assigned)

3.Limit of Detection (LoD)

Calibrator Concentration (pg/mL) RLU Mean Linear Equation
S1 0 8308 y=2514.4x+8308
S2 22.2 64128
Mean RLU values of zero - conc. diluent measured 20 times 8701
SD 427.7
Mean +2SD 9556
LoD (pg/mL) 0.496

Table 3. Limit of Detection of VB12

4.Specificity

Antigens (VB12, Adenosylcobalamin [AdoCbl], Hydroxocobalamin [HOCbl], and Methylcobalamin [MeCbl]) were diluted to 500 and 2000 pg/mL in 10mM PBS for detection.

Specificity of VB12 Antibody Recognition

Figure 3. Specificity of VB12 Antibody Recognition

Theoretical Conc.(pg/mL) Detected Conc.(pg/mL)
VB12 AdoCbl HOCbl MeCbl
0 0 0 0 0
500 499 487 705 656
2000 2015 1615 1943 2224

Table 4. Specificity of VB12 Antibody Recognition Data

5.Repeatability

The same batch of VB12 antibody (H835e1) and intrinsic factor (C1770) were used to detect Roche-assigned samples at concentrations of?50.26 pg/mL?and?523.7 pg/mL 10 times each. The measured concentrations exhibited?coefficients of variation (CV)?of?< 3%?for both low- and high-value samples.

Low-Concentration Samples
Sample RLU Concentration
(pg/mL)
1 107100 50.26
2 107866 50.87
3 107502 50.58
4 107213 50.35
5 107112 50.27
6 108017 50.99
7 106987 50.17
8 106547 49.82
9 104710 48.36
10 108306 51.22
Mean 107136 50.29
SD / 0.80
CV 1.6%

Table 5. VB12 Detection Repeatability
(Low-Concentration)

High-Concentration Samples
Sample RLU Concentration
(pg/mL)
1 991892 514.2
2 1010307 522.4
3 1003793 519.5
4 1037952 534.7
5 1009633 522.1
6 1074624 551.0
7 1041325 536.2
8 986505 511.8
9 1026037 529.4
10 1033005 532.5
Mean 1021507 527.4
SD / 11.79
CV 2.2%

Table 6. VB12 Detection Repeatability
(High-Concentration)

Free Sample Application

Free Sample Application

Item Introduction

Item Introduction

Vitamin B12 is the collective term for a number of substances with qualitatively equivalent biological effects. These compounds consist of a corrin ring system that is similar to porphyrin with four reduced pyrrole rings and a central cobalt ion and are therefore referred to as cobalamins. Characteristically, all cobalamins have an α-axial ligand at the cobalt ion consisting of a phosphoribosyl-5,6-dimethylbenzimidazole side chain. The β-axial position can be occupied by various substituents. Depending on the group (R), a distinction is made between cyano- (R = CN), aquo-(R = H2O), hydroxo- (R = OH), methyl- (R = CH3), and adenosyl-(R = 5’-deoxyadenosyl) cobalamin. Cobalamins are synthesized only by microorganisms, and humans and animals receive them solely through the food chain. In natural foods, vitamin B12 is present mainly as hydroxocobalamin and adenosylcobalamin and is found in milk as methylcobalamin. Due to its stability, cyanocobalamin is used in fortified foods as well as in pharmaceuticals and dietary supplements. Some pharmaceuticals and dietary supplements also contain hydroxocobalamin and methylcobalamin.

Vitamin B12 is necessary for the transformation of methyltetrahydrofolate to tetrahydrofolate for DNA synthesis. B12 deficiency can therefore lead to functional folate deficiency and impaired DNA synthesis, resulting in pernicious (megaloblastic) anaemia.

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