DAB, HRP, chromogenic, substrate, CAS, 91-95-2

FT-732320
DAB, HRP chromogenic insoluble substrate
Product description
A high quality DAB substrate solution for immunoblotting and immunohistochemical staining techniques
Name:
DAB solution HRP chromogenic substrate for IB and IHC
Product number:
UP732320, 500 ml
contains: 10ml DAB Substrate solution 50X (L)
50ml Buffer Stock Solution 10X
Storage:
Storage at +4°C.
Stable for 18 months at 2°C to 8°C.
Security note:
DAB is a suspected carcinogen.
H334
Introduction
DAB (3,3' Diaminobenzidine , CAS:91-95-2) is a widely used chromogenic substrate for Peroxidase (HRP) detections.
Our DAB solution contains 0.0694 M DAB of highest purity, and proprietary agents for enhancement of sensitivity and
stability. It is a 50x concentrate that is easily diluted with the provided buffer (supplied as 10x concentrate, pH 7.6 when
diluted) before use on immunoblots and slides that have been probes with HRP labelled probes.
Directions for use
Reagents
Tris/HCl Buffer 10X:
Dilute the provided 10X buffer with reagent grade water to 1X.
Reagent required but not provided:
Hydrogen peroxide 0,5%:
Dilute 1,67 ml of 3% hydrogen peroxide to 10 ml with reagent grade water.
Tris/Maleic acid buffer (for immunoblotting):
Dissolve 12,1 g of Tris base in approximately 800 ml of reagent grade water.
Adjust pH to 6,0 with a 2 Mol L-1 solution of maleic acid. Complete to 1L with reagent grade water.
Protocol 1 : DAB as HRP substrate in IHC technique
1- To 100 parts of Tris/HCl buffer, add 2 part of DAB concentrate and 1 part of 0,5% hydrogen peroxide.
Mix well and protect from light.
2- Completely cover tissue sections with this solution and incubate 5-15 min at room temperature.
3- After reaction is completed, wash sections thoroughly in reagent grade water.
4- Counterstain with hematoxylin if desired.
5-Dehydrate in graded alcohols to xylene or xylene substitutes.
6- Mount using xylene based mounting media.
Contact your local distributor
[email protected]
Uptima, powered by
P.1/2
FT-732320
Protocol 2 : DAB as HRP substrate in Blotting technique
1- To 100 parts of Tris/HCl buffer, add 2 part of DAB concentrate and 1 part of 0,5% hydrogen peroxide.
Mix well and protect from light.
2- Completely cover membranes with this solution and incubate 5-30 min at room temperature.
3- After reaction is completed, wash membranes thoroughly in reagent grade water.
4- Air dry and store protected from light.
Notes
1. Hydrogen peroxide is a powerful oxidizing agent; avoid skin contact.
2. DAB is a possible carcinogen; wear gloves when using it. The dry powder must never be used on the open bench; always use a fume cupboard.
3. It is possible that after use the copper sulphate solution in Step 18 may be contaminated with DAB; gloves should be worn when handling.
4. Do not allow the sections to dry out at any stage during the technique.
5. Dilute primary and biotinylated secondary antisera as appropriate in 1:20 normal swine serum.
CAUTION:
H334: Peut provoquer des symptômes allergiques ou d'asthme ou des difficultés respiratoires par inhalation
R36/37/38 : Peut irriter les yeux, le système respiratoire et la peau
R42/43 : Peut entrainer une sensibilisation par inhalation et contact avec la peau
S36/37/39
DAB is a suspected carcinogen ()
: Porter un vêtement de protection approprié, des gants et un appareil de protection des yeux/du visage
Related reagents
See BioSciences Innovations catalogue and e-search tool.
*other reagents for immunodetections with DAB substrate:
Hydrogen Peroxide #15983Q
WB:HP-labeled secondary antibodies (p.A324)
HRP labeled (strept)avidins (A350)
ProTran 0.2µm NC membranes, 20x20cm, #S31441
Blotting paper 1mm thick, 460x570cm, #BP2791
Stains: Protein Membrane Reversible stain #20078A
*Other substrates for HRP: chromogenic:
op-Metal Enhanced DAB kit #679921
DAB ultrapure powder #UP01012G
buffers and saturants (A365):
SeaBlock agent #UP40301A
TBS with non fat milk #GS4160
TBS with Tween20 #GS4200
BSA #UPQ84170 (powder) or #UP900130 (soln 30%)
BioBlock Saturating agent for blotting (inTBS) #N13650
Non fat milk powder #768701
chemiluminogenic:
UptiLight Classic #UP996190, spray #BM4961, UltraSensitive #58372A
Contact uptima – Interchim for any question
Contact your local distributor
[email protected]
Rev. R03E-I07vb
Uptima, powered by
P.2/2