Human Mcl-1 Antibody

Catalog #: AF8281 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF8281
AF8281-SP

Detection of Human Mcl‑1 by Western Blot.

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Human Mcl-1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Mcl-1 in direct ELISAs and Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Mcl-1
Val147-Gly219
Accession # Q07820

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.5 µg/mL

See below

Simple Western

20 µg/mL

Raji human Burkitt's lymphoma cell line.

Knockout Validated

Mcl‑1 is specifically detected in A431 human epithelial carcinoma parental cell line but is not detectable in Mcl‑1 knockout A431 cell line.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human Mcl‑1 by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line and Ramos human Burkitt's lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Mcl-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8281) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Mcl-1 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Knockout Validated

Western Blot Shows Human Mcl-1 Antibody Specificity by Using Knockout Cell Line.

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Western Blot Shows Human Mcl‑1 Specificity by Using Knockout Cell Line. Western blot shows lysates of A431 human epithelial carcinoma parental cell line and Mcl-1 knockout A431 cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Mcl-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8281) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Mcl-1 at approximately 40 kDa (as indicated) in the parental A431 cell line, but is not detectable in knockout A431 cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western

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Detection of Human Mcl‑1 by Simple Western^TM. Simple Western lane view shows lysates of Raji human Burkitt's lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Mcl‑1 at approximately 48 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human Mcl‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8281). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Mcl-1

MCL-1 (induced Myeloid Cell Leukemia differentiation protein 1; also Bcl2L3 and mcl1/EAT) is a 40-45 kDa member of the Bcl-2 family of proteins. It is widely expressed (in B cells, T cells, neutrophils and fibroblasts) and classified as a prosurvival Bcl-2 family member. Functionally, full-length MCL-1 sequesters the proapoptotic proteins Bak and Bax, rendering them inactive. It also delays cell-cycle progression by interacting with CDK1, CHK1 and PCNA. Human MCL-1 is a likely a 350 amino acid (aa) type II transmembrane protein. It contains a large cytoplasmic region (aa 1-327) plus a very short 2 aa C-terminal luminal segment. The cytoplasmic region has multiple domains, including a PEST (Pro/Glu/Ser/Thr)-like segment (aa 104-175), four ubiquitination sites, at least six utilized phosphorylation sites, and three Bcl2-like homology domains (aa 209-223; 252-272; 304-319). MCL-1 is known to form homodimers. There is one splice variant. It is 32-33 kDa in size and contains a 42 aa substitution for aa 230-350. This short form heterodimerizes with full-length MCL-1, rendering it incapable of interacting with Bak and Bax. MCL-1 also undergoes caspase processing. Cleavage after Asp127 generates 17 and 28-30 kDa fragments, while cleavage after Asp 157 generates 21 and 23-25 kDa fragments. These fragments give rise to a proapoptotic environment. Over aa 147-219, human MCL-1 shares 74% aa sequence identity with mouse Mcl-1.

Long Name

Myeloid Cell Leukemia Sequence 1

Entrez Gene IDs

4170 (Human); 17210 (Mouse)

Alternate Names

BCL2L3; bcl2-L-3; BCL2L3MGC104264; Bcl-2-like protein 3; Bcl-2-related protein EAT/mcl1; EAT; induced myeloid leukemia cell differentiation protein Mcl-1; Mcl1; Mcl-1; mcl1/EAT; MCL1-ES; MCL1L; MCL1S; MGC1839; myeloid cell leukemia ES; myeloid cell leukemia sequence 1 (BCL2-related); TM