alpha Actinin 4 Recombinant Antibody

$Blocking buffer: 5% NFDM\/TBST\r\nPrimary ab dilution: 1:5000\r\nPrimary ab incubation condition: 4\u00b0C overnight\r\nSecondary ab: Goat Anti-Rabbit IgG H&L (HRP)\r\nLysate: 1:HeLa, 2:HEK-293, 3:4T1, 4:Raw264.7\r\nProtein loading quantity: 20 \u03bcg\r\nExposure time: 30 s\r\nPredicted MW: 100 kDa\r\nObserved MW: 110 kDa\r\n$

Blocking buffer: 5% NFDM\/TBST\r\nPrimary ab dilution: 1:5000\r\nPrimary ab incubation condition: 4\u00b0C overnight\r\nSecondary ab: Goat Anti-Rabbit IgG H&L (HRP)\r\nLysate: 1:HeLa, 2:HEK-293, 3:4T1, 4:Raw264.7\r\nProtein loading quantity: 20 \u03bcg\r\nExposure time: 30 s\r\nPredicted MW: 100 kDa\r\nObserved MW: 110 kDa\r\n

$Tissue: Human colon Section type: Formalin fixed & Paraffin -embedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris\/EDTA buffer, pH 9.0 Primary ab dilution: 1:100 Primary ab incubation condition: 1 hour at room temperature Secondary ab: SP Kit(Rabbit) (sp-0023) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for bsm-60423R$

Tissue: Human colon Section type: Formalin fixed & Paraffin -embedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris\/EDTA buffer, pH 9.0 Primary ab dilution: 1:100 Primary ab incubation condition: 1 hour at room temperature Secondary ab: SP Kit(Rabbit) (sp-0023) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for bsm-60423R

$Cell line: HeLa\r\nFixative: 4% Paraformaldehyde\r\nPermeabilization: 0.1% TritonX-100\r\nPrimary ab dilution: 1:50\r\nPrimary incubation condition: 4\u00b0C overnight\r\nSecondary ab: Goat Anti-Rabbit IgG\r\nNuclear counter stain: DAPI (Blue)\r\nComment: Color green is the positive signal for bsm-60423R$

Cell line: HeLa\r\nFixative: 4% Paraformaldehyde\r\nPermeabilization: 0.1% TritonX-100\r\nPrimary ab dilution: 1:50\r\nPrimary incubation condition: 4\u00b0C overnight\r\nSecondary ab: Goat Anti-Rabbit IgG\r\nNuclear counter stain: DAPI (Blue)\r\nComment: Color green is the positive signal for bsm-60423R

$Lane 1: Mouse Skin tissue lysates; Lane 2: Mouse Uterus tissue lysates; Lane 3: Human HeLa cell lysates; Lane 4: Human Jurkat cell lysates; Lane 5: Human U251 cell lysates probed with alpha Actinin 4 Monoclonal Antibody, Unconjugated (bsm-60423R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.$

Lane 1: Mouse Skin tissue lysates; Lane 2: Mouse Uterus tissue lysates; Lane 3: Human HeLa cell lysates; Lane 4: Human Jurkat cell lysates; Lane 5: Human U251 cell lysates probed with alpha Actinin 4 Monoclonal Antibody, Unconjugated (bsm-60423R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Applications

WB
IHC-P
IHC-F
IF
ICC/IF

Reactivity

Human
Mouse

Overview
Catalog #	bsm-60423r
Product Name	alpha Actinin 4 Recombinant Antibody
Applications	WB, IHC-P, IHC-F, IF, ICC/IF
Reactivity	Human, Mouse
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic peptide derived from human alpha Actinin 4
Clonality	Recombinant
Isotype	IgG
Concentration	1mg/ml
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	81
Swiss Prot	O43707
Subcellular location	Cytoplasm, Nucleus
Synonyms	actinin 4; Actinin alpha 4; Actinin alpha-4; Alpha-actinin-4; _-Actinin-4; _Actinin-4; actinin4; ACTN 4; ACTN4; ACTN4_HUMAN; DKFZp686K23158; F actin cross linking protein; F-actin cross-linking protein; Focal segmental glomerulosclerosis 1; FSGS 1; FSGS; FSGS1; Non muscle alpha actinin 4; Non-muscle alpha-actinin 4.
Background	Alpha actinins belong to the spectrin gene superfamily which represents a diverse group of cytoskeletal proteins, including the alpha and beta spectrins and dystrophins. Alpha actinin is an actin-binding protein with multiple roles in different cell types. In nonmuscle cells, the cytoskeletal isoform is found along microfilament bundles and adherens-type junctions, where it is involved in binding actin to the membrane. In contrast, skeletal, cardiac, and smooth muscle isoforms are localized to the Z-disc and analogous dense bodies, where they help anchor the myofibrillar actin filaments. This gene encodes a nonmuscle, alpha actinin isoform which is concentrated in the cytoplasm, and thought to be involved in metastatic processes. Mutations in this gene have been associated with focal and segmental glomerulosclerosis. [provided by RefSeq, Jul 2008].
Application Dilution
WB	=1:500-2000
IHC-P	IHC-P=1:50-1:100
IHC-F	ICC=1:50
IF	IF=1:100-500
ICC/IF	1:50-100