IHC (formalin/paraffin): staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0 (30 min at RT). Use in ICC/IF was reported in the scientific literature (PMID: 23813946).
Caspase 3 is a member of the interleukin-1 BETA-converting enzyme family. Caspase 3 is thought to be associated with induction of apoptosis.Caspase 3 is ubiquitously expressed and like other caspases is synthesized as an inactive (32kDa) proenzyme. Upon activation, Caspase 3 is cleaved at Asp28-Ser29 and Asp175-Ser176 thereby generating two subunits of 17kDa and 12kDa, respectively. Recent studies have implicated that Caspase 3 is associated with the induction of apoptosis. Activation of Caspase 3 occurs in response to variety of apoptotic inducers including Fas mediated apoptosis.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
FAQs for Caspase-3 Antibody (NB600-1235). (Showing 1 - 4 of 4 FAQs).
Any reason for cleaved caspase 3 to be seen as purely nuclear staining or is this just an artifact?
Caspase 3 can translocate from the cytoplasm to the nucleus, usually during apoptosis. We do offer a 100% guarantee that this antibody will be useful as stated on the datasheet also.
The datasheet says that it reacts with the active form of Caspase 3 (17 kDa protein), but every time when I do the WB, I get two bands between 15kDa and 20kDa. The different company's cleaved caspase 3 antibody reacts with 17 and 19 kDa protein, so I wonder which band is specific to yours.
Regarding your question, NB600-1235 has been exclusively validated for IHC, not western blot and it is supposed to detect the active form of caspase3. The inactive Caspase 3 is a 31 kD protein, a heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 17 kDa (p17) and a 12 kDa (p12). When it is Cleaved by granzyme B, it generates the two active subunits. As a result the two bands you are observing on the gel should be correct. However, we haven't tested this antibody for western blot.
I'd like to get the information about the active/cleaved Caspase 3 Antibody (NB600-1235). The datasheet says that it reacts with the active form of Caspase 3 (17 kDa protein), but every time when I do the WB, I get two bands between 15kDa and 20kDa. The different company's cleaved caspase 3 antibody reacts with 17 and 19 kDa protein, so I wonder which band is specific to yours?
According to the data I could find for the active/cleaved caspase 3, the active form should appear at approximately 17kDa and also has a smaller subunit seen at approximately 12kDa. The inactive form is approximately 32kDa. These are the theoretical weights and can vary based on type of gel and samples they are run on. In most cases, it looks like most antibodies detect the active forms at 19 and 17kDa respectively. With this particular product, NB600-1235, we have not yet validated it for use in western blot, so we unfortunately do not have any information for the location of bands.
This is a Rabbit antibody right so Do you think I can use it on rabbit tissue?
This antibody does react to rabbit but if you are using it with an anti-rabbit secondary, you will almost certainly see some background staining. I know that mouse-on-mouse blocking systems exist, but I am not sure if there is a rabbit equivalent at this time. The best way to use NB600-1235 on rabbit tissues would be to have it directly conjugated to biotin, HRP, or whatever detection method you are planning to use. This will eliminate the need for an anti-rabbit secondary and should give you little to no background. We do offer a conjugation service, so if you would like to have this antibody conjugated, please let me know what you would like to have this antibody conjugated to and I can get you a quote, email firstname.lastname@example.org.
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