MCL1 Recombinant monoclonal antibody

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  WB analysis using 82695-4-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 82695-4-RR (MCL1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IHC staining of human intrahepatic cholangiocarcinoma using 82695-4-RR

  Immunohistochemical analysis of paraffin-embedded human intrahepatic cholangiocarcinoma tissue slide using 82695-4-RR (MCL1 antibody) at dilution of 1:1000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human intrahepatic cholangiocarcinoma using 82695-4-RR

  Immunohistochemical analysis of paraffin-embedded human intrahepatic cholangiocarcinoma tissue slide using 82695-4-RR (MCL1 antibody) at dilution of 1:1000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human ovarian cancer using 82695-4-RR

  Immunohistochemical analysis of paraffin-embedded human ovarian cancer slide using 82695-4-RR (MCL1 antibody) at dilution of 1:200 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human ovarian cancer using 82695-4-RR

  Immunohistochemical analysis of paraffin-embedded human ovarian cancer slide using 82695-4-RR (MCL1 antibody) at dilution of 1:200 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of MCF-7 using 82695-4-RR

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using MCL1 antibody (82695-4-RR, Clone: 5P13 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  FC experiment of Ramos using 82695-4-RR

  1x10^6 Ramos cells were intracellularly stained with 0.25 ug MCL1 Recombinant antibody (82695-4-RR, Clone:5P13) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG Isotype Control Recombinant Antibody (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer.

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  Affinity and Kinetic Characterization of 82695-4-RR

  Biolayer interferometry (BLl) kinetic assays of 82695-4-RR against Human MCL1 were performed. The affinity constant is 2.13 nM.

MCL1 Uni-rAb^® Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Cat No. 82695-4-RR

Clone No.5P13

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Synonyms

5P13, Bcl2-L-3, Bcl-2-like protein 3, Bcl-2-related protein EAT/mcl1, EAT

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, MDA-MB-231 cells, Jurkat cells, Raji cells
• Positive IHC detected in: human intrahepatic cholangiocarcinoma tissue, human ovarian cancer; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: MCF-7 cells
• Positive FC (Intra) detected in: Ramos cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:500-1:2000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

82695-4-RR targets MCL1 in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen:

  CatNo: Ag10397

  Product name: Recombinant human MCL1 protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 1-327 aa of BC107735

  Sequence: MFGLKRNAVIGLNLYCGGAGLGAGSGGATRPGGRLLATEKEASARREIGGGEAGAVIGGSAGASPPSTLTPDSRRVARPPPIGAEVPDVTATPARLLFFAPTRRAAPLEEMEAPAADAIMSPEEELDGYEPEPLGKRPAVLPLLELVGESGNNTSTDGSLPSTPPPAEEEEDELYRQSLEIISRYLREQATGAKDTKPMGRSGATSRKALETLRRVGDGVQRNHETAFQGMLRKLDIKNEDDVKSLSRVMIHVFSDGVTNWGRIVTLISFGAFVAKHLKTINQESCIEPLAESITDVLVRTKRDWLVKQRGWDGFVEFFHVEDLEGG

• Full Name: myeloid cell leukemia sequence 1 (BCL2-related)
• Calculated Molecular Weight: 350 aa, 37 kDa
• Observed Molecular Weight: 40 kDa
• GenBank Accession Number: BC107735
• Gene Symbol: MCL1
• Gene ID (NCBI): 4170
• RRID: AB_3086509
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: Q07820
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

MCL-1 is an anti-apoptotic member of the Bcl-2 family originally isolated from the ML-1 human myeloid leukemia cell line. Similar to BCL2 and BCL2L1, MCL1 can interact with BAX and/or BAK1 to inhibit mitochondria-mediated apoptosis. Mcl-1 is critical for the proliferation and survival of myeloma cells in vitro, and overexpression of Mcl-1 protein in myeloma cells is associated with relapse and short event-free survival in multiple myeloma patients. Recent studies show that MCL-1 is upregulated in numerous haematological and solid tumor malignancies. Therefore, MCL-1 has been suggested as a potential new therapeutic target. MCL-1 can be alternatively spliced into a long form (MCL-1L, 40 kDa) or a short form (MCL-1S, 30 kDa). (PMID: 15467463, PMID: 15147382)