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Recombinant Rabbit Monoclonal
Anti-Vimentin Rabbit mAb
Catalog Number: PTM-5376
$ 280

Clone Number: JRMR-10214-73

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P ICC/IF FC IP mIHC-P

Reactivity: Human, Mouse, Rat

Synonyms: VIM

Product Size
100 μl
Quantity

Shipping: Ambient temperature

Order online or send purchase order to info@ptmbio.com

FAQ Technical Support Protocols

General Information
Isotype IgG
Conjugate Unconjugated
Synonyms VIM
UniProt ID

P08670

Immunogen Recombinant human Vimentin fragment
MW (kDa) 54
Specificity Anti-Vimentin Rabbit mAb recognizes endogeneous level of Vimentin protein.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:100 - 1:500 Human, Mouse, Rat
ICC/IF 1:50 Human, Mouse
FC 1:50 - 1:100 Human
IP 1:25 - 1:100 Human
mIHC-P 1:500 - 1:1000 Human
Properties
Purity Protein A purified
Constituents PBS, Glycerol, BSA
Storage Store at -20°C. Avoid freeze/thaw cycles.
Stability Stable for 12 months from date of receipt/reconstitution.
Target Information

Background

Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally. Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2.

Cellular location

Cytoskeleton, Cytoplasm

Images
WB

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Lysate: HeLa, HEK-293, Jurkat, Rat brain, Mouse brain
Protein loading quantity: 20 μg
Exposure time: 3 s
Predicted band size: 54 kDa
Observed band size: 54 kDa
IHC-P

Tissue: Human cervical carcinoma
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: 1:500
Primary Ab incubation: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter stain: Hematoxylin (Blue)
Description: The brown color represents the positive signal observed with PTM-5376

Tissue: Mouse kidney
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: 1:500
Primary Ab incubation: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter stain: Hematoxylin (Blue)
Description: The brown color represents the positive signal observed with PTM-5376

Tissue: Rat kidney
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: 1:500
Primary Ab incubation: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter stain: Hematoxylin (Blue)
Description: The brown color represents the positive signal observed with PTM-5376
ICC/IF

Cell line: HeLa
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% TritonX-100
Primary Ab dilution: 1:50
Primary incubation condition: 4°C overnight
Secondary Ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (Blue)
Description: The green color represents the positive signal observed with PTM-5376

Cell line: NIH/3T3
Fixative: 100% Ice-cold methanol
Permeabilization: 0.1% TritonX-100
Primary Ab dilution: 1:50
Primary incubation condition: 4°C overnight
Secondary Ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (Blue)
Description: The green color represents the positive signal observed with PTM-5376
FC

Cell line: HeLa
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% TritonX-100
Primary Ab dilution: 1:100
Secondary Ab: Goat anti Rabbit IgG
Unlabeled control: Cells without incubation with primary and secondary antibodies (Black line).
Isotype control: Rabbit monoclonal IgG (Blue line).
Description: Line red is the positive signal for PTM-5376
IP

IP of HeLa cells extracts
IP Ab incubation: PTM-5376, 4°C overnight, 1:100 dilution
WB Primary Ab incubation: PTM5376, room temperature 2h, 1:2000 dilution
Secondary Ab: Anti-Rabbit IgG for IP (HRP)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
1: 5% Input
2: IP with Rabbit monoclonal IgG Isotype
3: IP with PTM-5376
Observed MW: 54 kDa
Exposure time: 30 s
mIHC-P

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. Heat mediated antigen retrieval (Tris/EDTA buffer, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

Tissue: Human stomach
Section type: Formalin fixed & Paraffin - embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0. Heat mediated antigen retrieval (Tris/EDTA buffer, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.
Primary ab dilution: PTM-5376 (1:1000), PTM-6334 (1:100), PTM-6420 (1:1000)
Primary Ab incubation: 90 minutes at room temperature
Secondary ab: Anti-Rabbit and Mouse Polymer HRP (Ready to use)
Nuclear counter stain: DAPI (Blue)
Comment: Color red is the positive signal for Mucin-6 (PTM-6420, TSA 570), color green is the positive signal for Vimentin ( PTM-5376, TSA 520), color yellow is the positive signal for Mucin 5AC ( PTM-6334, TSA 650). The section was incubated in three rounds of staining; in the order of PTM-5376,PTM-6334,PTM-6420; each using a separate fluorescent tyramide signal amplification system.
Primary ab dilution: PTM-5376 (1:1000), PTM-5995 (1:500)
Primary Ab incubation: 90 minutes at room temperature
Secondary ab: Anti-Rabbit and Mouse Polymer HRP (Ready to use)
Nuclear counter stain: DAPI (Blue)
Comment: Color red is the positive signal for MUC-2 (PTM-5995, TSA 570), color green is the positive signal for Vimentin ( PTM-5376, TSA 520). The section was incubated in two rounds of staining; in the order of PTM-5376, PTM-5995; each using a separate fluorescent tyramide signal amplification system.

Research Use

For research use only, not for use in diagnostic procedures.

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