- MA3-903 - Invitrogen Antibodies HCN4 antibody

Cain SM, Tyson JR, Jones KL, Snutch TP
Pflugers Archiv : European journal of physiology 2015 Jun;467(6):1367-82

Müller F, Scholten A, Ivanova E, Haverkamp S, Kremmer E, Kaupp UB
The European journal of neuroscience 2003 May;17(10):2084-96

Stevens DR, Seifert R, Bufe B, Müller F, Kremmer E, Gauss R, Meyerhof W, Kaupp UB, Lindemann B
Nature 2001 Oct 11;413(6856):631-5

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HCN4 was performed by loading 25 µg of rat eye lysate onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a HCN4 monoclonal antibody (Product # MA3-903) at a dilution of 1:200 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~132 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal deparaffinized Human heart 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 (Product # 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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 (Product # 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 HCN protein isoforms are differentially expressed in GAERS VB thalamus. a Representative Western blots for HCN proteins from adult NEC and GAERS VB samples. Primary antibodies to detect each of the HCN channel isoforms were compared to vinculin in order to normalize the expression data. b Mean Western blot densitometry data for HCN1-4 normalized to vinculin for NEC ( n = 12 samples (six animals); black columns ) and GAERS ( n = 12 samples (six animals); grey columns ) VB samples

MA3-903