Human B-cell lymphoma/leukemia 10 (BCL10) ELISA Kit (HUEB2680)

Product Name:	Human B-cell lymphoma/leukemia 10 (BCL10) ELISA Kit
SKU:	HUEB2680
Size:	96T
Target:	Human B-cell lymphoma/leukemia 10 (BCL10)
Synonyms:	B-cell CLL/lymphoma 10, CARD-containing molecule enhancing NF-kappa-B, CARD-like apoptotic protein, CED-3/ICH-1 prodomain homologous E10-like regulator, Cellular homolog of vCARMEN, Cellular-E10, Mammalian CARD-containing adapter molecule E10, Bcl-10, hCLAP, CIPER, cCARMEN, c-E10, mE10, CIPER, CLAP
Assay Type:	Sandwich
Detection Method:	ELISA
Reactivity:	Human
Detection Range:	78-5000pg/mL
Sensitivity:	39.4pg/mL

Intra CV:

7.2%

Inter CV:

8.5%

Linearity:

Sample	1:2	1:4	1:8	1:16
Serum(N=5)	104-117%	106-115%	100-110%	99-111%
EDTA Plasma(N=5)	81-94%	102-112%	105-115%	110-120%
Heparin Plasma(N=5)	99-109%	98-108%	100-112%	92-101%

Recovery:

Sample Type	Average(%)	Recovery Range(%)
Serum	107	101-113
Plasma	109	103-115

Function:

Involved in adaptive immune response (PubMed:25365219). Promotes apoptosis, pro-caspase-9 maturation and activation of NF-kappa-B via NIK and IKK. May be an adapter protein between upstream TNFR1-TRADD-RIP complex and the downstream NIK-IKK-IKAP complex. Is a substrate for MALT1 (PubMed:18264101).

Uniprot:	O95999
Sample Type:	Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:	Natural and recombinant human B-cell lymphoma/leukemia 10
Sub Unit:	Found in a membrane raft complex, at least composed of BCL10, CARD11, DPP4 and IKBKB. Self-associates by CARD-CARD interaction and forms a tight complex with MALT1. Interacts with other CARD-proteins such as CARD9, CARD10, CARD11 and CARD14. Binds caspase-9 with its C-terminal domain. Interacts with TRAF2 and BIRC2/c-IAP2. Interacts with PELI2 and SOCS3; these interactions may be mutually exclusive.
Research Area:	Cancer
Subcellular Location:	Cytoplasm Perinuclear region Membrane raft Appears to have a perinuclear, compact and filamentous pattern of expression. Also found in the nucleus of several types of tumor cells. Colocalized with DPP4 in membrane rafts.
Storage:	Please see kit components below for exact storage details
Note:	For research use only

UniProt Protein Function:	Bcl-10: a ubiquitous pro-apoptotic protein that promotes pro-caspase-9 maturation and activation of NF-kappa-B via NIK and IKK. May be an adapter protein between upstream TNFR1-TRADD-RIP complex and the downstream NIK-IKK-IKAP complex. Self-associates via CARD-CARD interactions; forms a tight complex with MALT1. Interacts with other CARD-proteins such as CARD9, CARD10, CARD11 and CARD14. Binds caspase-9 with its C-terminal domain. Interacts with TRAF2 and BIRC2/c-IAP2. Defects in BCL10 are involved in various types of cancer.
UniProt Protein Details:	Protein type:Apoptosis; Tumor suppressor; Transcription, coactivator/corepressor Chromosomal Location of Human Ortholog: 1p22 Cellular Component: CBM complex; protein complex; lysosome; immunological synapse; cytosol; lipid raft; T cell receptor complex; cytoplasmic microtubule; perinuclear region of cytoplasm; cytoplasm; plasma membrane; lipopolysaccharide receptor complex; nucleus Molecular Function:protein C-terminus binding; protein kinase B binding; ubiquitin binding; protease binding; protein self-association; transcription coactivator activity; transcription factor binding; protein kinase binding; NF-kappaB binding; protein binding; enzyme binding; ubiquitin protein ligase binding; kinase activator activity; kinase binding Biological Process: cell death; response to fungus; positive regulation of transcription, DNA-dependent; positive regulation of caspase activity; B cell apoptosis; T cell receptor signaling pathway; interleukin-6 biosynthetic process; activation of NF-kappaB transcription factor; negative regulation of mature B cell apoptosis; response to molecule of bacterial origin; protein homooligomerization; response to food; adaptive immune response; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of interleukin-8 biosynthetic process; protein oligomerization; positive regulation of protein ubiquitination; lymphotoxin A biosynthetic process; regulation of T cell receptor signaling pathway; neural tube closure; toll-like receptor signaling pathway; innate immune response; cellular defense response; positive regulation of mast cell cytokine production; positive regulation of T cell activation; immunoglobulin mediated immune response; positive regulation of phosphorylation Disease: Follicular Lymphoma, Susceptibility To, 1; Mesothelioma, Malignant; Gastric Lymphoma, Primary; Testicular Germ Cell Tumor; Immunodeficiency 37
NCBI Summary:	This gene was identified by its translocation in a case of mucosa-associated lymphoid tissue (MALT) lymphoma. The protein encoded by this gene contains a caspase recruitment domain (CARD), and has been shown to induce apoptosis and to activate NF-kappaB. This protein is reported to interact with other CARD domain containing proteins including CARD9, 10, 11 and 14, which are thought to function as upstream regulators in NF-kappaB signaling. This protein is found to form a complex with MALT1, a protein encoded by another gene known to be translocated in MALT lymphoma. MALT1 and this protein are thought to synergize in the activation of NF-kappaB, and the deregulation of either of them may contribute to the same pathogenetic process that leads to the malignancy. [provided by RefSeq, Jul 2008]
UniProt Code:	O95999
NCBI GenInfo Identifier:	4502379
NCBI Gene ID:	8915
NCBI Accession:	NP_003912.1
UniProt Secondary Accession:	O95999,Q5VUF1,
UniProt Related Accession:	O95999
Molecular Weight:	26,252 Da
NCBI Full Name:	B-cell lymphoma/leukemia 10
NCBI Synonym Full Names:	B-cell CLL/lymphoma 10
NCBI Official Symbol:	BCL10
NCBI Official Synonym Symbols:	CLAP; mE10; CIPER; c-E10; CARMEN
NCBI Protein Information:	B-cell lymphoma/leukemia 10; hCLAP; cCARMEN; cellular-E10; cellular homolog of vCARMEN; CARD-containing proapoptotic protein; CARD containing molecule enhancing NF-kB; CARD-containing apoptotic signaling protein; caspase-recruiting domain-containing prote
UniProt Protein Name:	B-cell lymphoma/leukemia 10
UniProt Synonym Protein Names:	B-cell CLL/lymphoma 10; Bcl-10; CARD-containing molecule enhancing NF-kappa-B; CARD-like apoptotic protein; hCLAP; CED-3/ICH-1 prodomain homologous E10-like regulator; CIPER; Cellular homolog of vCARMEN; cCARMEN; Cellular-E10; c-E10; Mammalian CARD-containing adapter molecule E10
Protein Family:	B-cell lymphoma/leukemia
UniProt Gene Name:	BCL10
UniProt Entry Name:	BCL10_HUMAN

Component	Quantity (96 Assays)	Storage
ELISA Microplate (Dismountable)	8×12 strips	-20°C
Lyophilized Standard	2	-20°C
Sample Diluent	20ml	-20°C
Assay Diluent A	10mL	-20°C
Assay Diluent B	10mL	-20°C
Detection Reagent A	120µL	-20°C
Detection Reagent B	120µL	-20°C
Wash Buffer	30mL	4°C
Substrate	10mL	4°C
Stop Solution	10mL	4°C
Plate Sealer	5	-

Other materials and equipment required:

Microplate reader with 450 nm wavelength filter
Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
Incubator
Deionized or distilled water
Absorbent paper
Buffer resevoir

*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1.	Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2.	Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3.	Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4.	Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5.	Repeat the wash process for five times as conducted in step 3.
6.	Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7.	Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8.	Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9.	After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type	Protocol
Serum	If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
Plasma	Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid	Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatant	Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysates	Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenates	The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysates	Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk	Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

ELISA	Antibodies
Human Bcl-10 (B-cell Leukemia/Lymphoma 10) ELISA Kit (HUES01741)	Anti-Bcl10 Antibody (CAB1106)[KO Validated]
	Anti-Bcl10 Antibody (CAB4520)
	EIF2S2 Antibody (PACO06000)
	BCCIP Antibody (PACO07989)
	BCAT2 Antibody (PACO15856)

Human B-cell lymphoma/leukemia 10 (BCL10) ELISA Kit (HUEB2680)

Description