Immunohistochemistry was performed on normal deparaffinized Human tonsil tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rat monoclonal antibody recognizing HCN4 (MA3-903) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat|
|Host / Isotype||Rat / IgG1|
|Immunogen||Synthetic peptide sequence corresponding to residues S H G S L L L P P A S S P P P P Q V P Q R R G T P P L T P G R L T Q D L K L of HCN4.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1/50|
|Immunofluorescence (IF)||Assay Dependent|
|Immunohistochemistry (Frozen) (IHC (F))||1:1000|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
MA3-903 detects HCN4 from human, mouse, and rat samples.
MA3-903 has been successfully used in Western blotting and immunohistochemistry procedures. By Western blot MA3-903 detects a ~132 KDa band representing HCN4.
The MA3-903 immunogen is a synthetic peptide sequence corresponding to residues S H G S L L L P P A S S P P P P Q V P Q R R G T P P L T P G R L T Q D L K L of HCN4.
MA3-903 clone SHG 1E5 is a rat monoclonal hybridoma. This antibody was produced by immunizing a rat, isolating the spleen cells, creating the hybridoma and successively injecting these cells into a mouse to produce ascites.
HCN4 is a member of the family of hyperpolarization-activated and cyclic-nucleotide-gated (HCN) channels. HCN currents have been linked to pacemaker activity in the heart and brain, resting potential control, as well as neuronal plasticity. It has been shown that HCN4 channels function as receptors for sour taste, and are associated with pacemaker potential generation in the sinoatrial node.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Thalamocortical neurons display suppressed burst-firing due to an enhanced Ih current in a genetic model of absence epilepsy.
MA3-903 was used in western blot to study burst-firing in TR neurons and spike-and-wave discharges in the Genetic Absence Epilepsy Rats from Strasbourg model
|Cain SM,Tyson JR,Jones KL,Snutch TP||Pflugers Archiv : European journal of physiology (467:1367)||2015|