We are looking into protein kinase C-alpha antibodies for immunohistochemistry on formalin fixed, paraffin embedded tissue. I see that you have many antibodies, including several monoclonals. I wanted to confirm with you that the following antibodies are specific for C-alpha. NB600-201=MC5 (an older paper suggested that this might react with alpha, beta, gamm), NB110-57356=Y124, NB110-57353=Y143. Also, if you have an comparator data or feedback regarding PKC-alpha antibodies for IHC on FFPE, that would be very helpful. We would like to pick 1 antibody that is easy to titer and robust, so that we can move forward with our studies.

Name: PKC alpha Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-04234
Price: 679 USD

Question

We are looking into protein kinase C-alpha antibodies for immunohistochemistry on formalin fixed, paraffin embedded tissue.  I see that you have many antibodies, including several monoclonals.  I wanted to confirm with you that the following antibodies are specific for C-alpha.  NB600-201=MC5 (an older paper suggested that this might react with alpha, beta, gamm), NB110-57356=Y124, NB110-57353=Y143.  Also, if you have an comparator data or feedback regarding PKC-alpha antibodies for IHC on FFPE, that would be very helpful. We would like to pick 1 antibody that is easy to titer and robust, so that we can move forward with our studies.

Accepted Answer

We have never heard any feedback that these cross-react with other PKC proteins. If you have seen data that MC5 may cross-react, then I would consider avoiding this antibody if you are concerned. The immunogens for NB110-57356 and NB110-57353 come from the N- and C-terminals, which is where the PKC proteins vary the most, so these are not expected to cross-react and we have had no feedback or testing data that suggests that they do. I would recommend NB110-57356, since it has been published with. You may want to review the published data to determine if it seems this antibody will suit your needs.

At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Reactivity	HuSpecies Glossary
Applications	WB

Dilutions	Western Blot
Application Notes	This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Theoretical MW	76.6 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Storage	Store at -80C. Avoid freeze-thaw cycles.
Buffer	RIPA buffer

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