Literature references (previous batch)
:
Wickstrom C, Davies JR, Eriksen GV, Veerman EC, Carlstedt I. MUC5B
is a major gel-forming, oligomeric mucin from human salivary gland, respiratory
tract and endocervix: identification of glycoforms and C-terminal cleavage.
Biochem J. 1998 Sep 15;334 ( Pt 3):685-93
Origin, source : Rabbits were immunised with a synthetic peptide coupled with a carrier, the KLH (Keyhole limpet hemacyanin). The peptide of 12 amino acid (RNREQVGKF KMC) is located in 4 of the cysteine domains of the tandem repeat region.
Specificity : Lum5B-2 recognizes the unreduced MUC5B glycoprotein but a stronger signal is obtained after reduction
Cross-reactivity : Lum5B-2 does not react with preparations of the mucin MUC2 (either unreduced or reduced) or MUC6 but a weak signal with a MUC5AC preparation was obtained in London on ELISA plates. No cross reactivity was observed with blood group substances ( A, LeA, LeB, H) obtained from ovarian cyst fluid.
Binding Assays : The Lum5B-2 antibody reacts with salivary mucin and respiratory cervical mucin purified by two consecutive density gradient centrifugations. The experiments were performed after reduction and alkylation of the samples.
Immunoblotting : Lum5B-2 antibody recognises the gel fraction (reduced or unreduced) and the soluble fraction (reduced) of induced sputum.
Results of immunohistochemistry experiments
performed in Ingemar Calstedt's lab :
Tissues fixed in formol/saline. Treatment : Sodium citrate
pH 6.
| Positive tissues: |
| Cervical canal (surface epithelium, glands) |
| Submandibular salivary gland (few focal points in the tissue) |
| Sublingual gland (secretoty acini) |
| Trachea (submucosal glands, epithelial goblet cells) |
Results of immunohistochemistry experiments
performed in Carme De Bolos's lab :
Treatment: Pressure cooker in citrate buffer
| Positive tissues: | Negative tissues: |
| Salivary gland (3/3)
Stomach (8/8) Jejunum (3/3) Colon (5/5) Gallbladder (1/1) Pancreas (4/4) Liver (1/3) Bronchial epithelium (3/3) Endocervix (4/4) Ovary (3/3) |
Oesophagus (0/3)
Duodenum (0/2) Ileum (0/2) Epydidimis (0/2) Anal mucosa (0/1) Vas deferens (0/2) Kidney (0/3) Seminal vesicle (0/1) Bladder (0/1) Prostate (0/1) Breast (0/1) Thyroid gland (0/2) Fallopian tube (0/1) Parathyroid gland (0/2) Testis (0/3) Exocervix (0/5) |
Results of immunohistochemistry experiments
performed in Jean-Pierre Aubert's lab :
Fixation in neutral buffered formalin
Treatment: microwave pretreatment in citrate buffer pH 6
| Positive tissues: | Negative tissues: |
| Bronchial epithelium (some goblet cells, gland
ducts and mucus glands)
Gall Bladder (surface epithelium) HT29 cells in culture(5% positive) |
Small intestine |
Results of immunohistochemistry experiments performed in Leonor David's lab :
Positive reactivity in the serous cells (cytoplasm) of the salivary
glands.
Results of immunohistochemistry and experiments performed in Jacques Bara's lab :
The Lum5B-2 polyclonal antibody immunoreacts in the same way as mMUC5B-2
EU1 and mMUC5B-2 EU2 monoclonal antibodies.
| Tissues | Immunohistology | ISH | |
| Oesophageal glands | +++ | ND | |
| Cardia | Surface
Glands |
-
- |
ND
ND |
| Fundus | Surface
Glands |
-
- |
-
- |
| Pylorus | Surface
Glands |
-
- |
-
- |
| Duodenum | Villi
Brunner's glands |
-
- |
-
- |
| Jejunum | Villi
Glands |
-
- |
-
- |
| Ileum | Villi
Glands |
-
- |
-
- |
| Colon | + | +* | |
| Prostate | (utriculus) | + | ??? |
| Conjunctiva | - | - | |
| Endocervix | ++++ | + | |
| Respiratory tract | Surface
Glands mucus cells Serous cells |
+++++
+++ - |
-
+++ - |
| Bile duct | +++ | + | |
| Pancreas | Islets of Langherans | - | - |
| Pancreas | Wirsung duct
Small ducts |
ND
+++ |
ND
+ |
| Gall Bladder | +++ | +++++ |
* B. Jan-Willem Van Klinken et al. Am. J. Physiol. 274 (37) G871 (1998)
More results from Jacques Bara's lab:
| Positives tissues: | Negatives tissues: |
| Caecum (Goblet cells-very slight staining)
Right colon (Goblet cells-very slight staining) Transverse colon (Goblet cells-very slight staining) Left colon (Goblet cells-moderate staining) Liver (Duct- moderate staining) Oviduct (mucus cells-slight staining) |
Oesophagus
Caecum (Abspt cells) Right colon (Abspt cells) Transverse colon (Abspt cells) Left colon (Abspt cells) Liver (Hepatocyte) Lung Ovary Bladder Adrenal glands |
Other literature references using the previous
batch of the antibody :
Davies JR, Svitacheva N, Lannefors L, Kornfalt R, Carlstedt I. Identification
of MUC5B, MUC5AC and small amounts of MUC2 mucins in cystic fibrosis airway
secretions.
Biochem J. 1999 Dec 1;344 Pt 2:321-30.
Note : Made for the EU Consortium