Vimentin Recombinant Rabbit Monoclonal Antibody

货号：

AWA10146

应用：

WB,IHC-P,IHC-F,IF-C,IF-T,IP,mIHC,FCM

反应性：

Human,Mouse,Rat,Monkey,Pig

来源：

Rabbit

规格 价格 库存
20μL ¥620 1-3个工作日
50μL ¥1250 1-3个工作日
100μL ¥2200 1-3个工作日

产品概述

Product Details

Host Species:

Rabbit

Reactivity:

Human, Mouse, Rat, Monkey, Pig

Molecular Wt:

Predicted MW: 54 kDa
Observed MW: 54 kDa

Clonality:

Monoclonal

Isotype:

IgG

Concentration:

1.185mg/ml

Other Names:

VIM; CTRCT30; Epididymis luminal protein 113; FLJ36605; HEL113; Vimentin

Formulation:

Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide.

Purification:

Affinity-chromatography

Storage:

Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20°C storage.

Applications

WB 1:1000-1:10000
IHC-P 1:100-1:1000
IHC-F 1:100-1:1000
IF-C 1:50-1:500
IF-T 1:100-1:1000
IP 1:100-1:300
mIHC 1:100-1:1000
FCM 1:50-1:200

Immunogen
Information

Gene Name:

VIM

Protein Name:

Vimentin

Gene ID:

7431 (Human)
22352 (Mouse)
81818 (Rat)

SwissPro:

P08670 (Human)
P20152 (Mouse)
P31000 (Rat)

Immunogen
Information

Subcellular Location:

Cytoplasm. Cytoplasm, cytoskeleton. Nucleus matrix. Cell membrane.

Immunogen:

Synthetic peptide within C-terminal human Vimentin.

Specificity:

Vimentin Monoclonal Antibody detects endogenous levels of Vimentin protein.

Product images
	Fig: Multiplex immunohistochemistry analysis of Rat-kidney tissue (Formalin/PFA-fixed paraffin-embedded sections). Merged staining of anti-LDHA (AWA10478; green; TSA-520:AWI0688), anti-vimentin (AWA10146; red; TSA-570: AWI0689), anti-Aquaporin 4 (AWA10201; white; TSA-690: AWI0691).Antibody Wash Solution (for mIHC) (AWI0707, 37℃, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. The section was incubated in three rounds of staining; in the order of LDHA (AWA10478) (1/200 dilution), anti-vimentin (AWA10146) (1/400 dilution), anti-AQP4 (AWA10201) (1/200 dilution); each using a separate fluorescent tyramide signal amplification system. DAPI (blue, AWC0291) was used as a nuclear counter stain. Microscopy and pseudocoloring of individual TSA dyes was performed using a pannoramic midi.

	Fig :Western blot analysis of Vimentin on different lysates. Proteins were transferred to a NC membrane and blocked with 5% NF-Milk in TBST for 1 hour at room temperature. The primary antibody (AWA10146, 1/1000) was used in TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (AWS0002) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control:* Lane 1: SK-N-SH cell Lane 2: A204 cell Lane 3: NIH3T3 cell Lane 4: L929 cell Lane 5: GL261 cell Lane 6: 3D4/21 cell Predicted molecular weight: 54 kDa Observed molecular weight: 54 kDa

	Fig : Western blot analysis of Vimentin on different lysates. Proteins were transferred to a NC membrane and blocked with 5% NF-Milk in TBST for 1 hour at room temperature. The primary antibody (AWA10146, 1/1000) was used in TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (AWS0002) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control:* Lane 1: A549 cell Lane 2: L6 cell Lane 3: RAW264.7 cell Lane 4: Hela cell Lane 5: HUVEC cell Lane 6: Jurkat cell Lane 7: COS7 cell Predicted molecular weight:54 kDa Observed molecular weight:54 kDa

	Fig: Multiplex immunohistochemistry analysis of Rat-Myocardial muscle tissue (Formalin/PFA-fixed paraffin-embedded sections). Merged staining of anti-Cardiac Troponin T (AWA10274; green; TSA-520:AWI0688), anti-Vimentin (AWA10146; red; TSA-570: AWI0689), anti-SM22 (AWA10249; white; TSA-690: AWI0691). Antibody Wash Solution (for mIHC) (AWI0707, 37℃, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. The section was incubated in three rounds of staining; in the order of Cardiac Troponin T (AWA10274) (1/200 dilution), anti-Vimentin (AWA10146) (1/600 dilution), anti-SM22 (AWA10249) (1/1000 dilution); each using a separate fluorescent tyramide signal amplification system. DAPI (blue, AWC0291) was used as a nuclear counter stain. Microscopy and pseudocoloring of individual TSA dyes was performed using a pannoramic midi.

	Fig: Immunocytochemistry analysis of HELA cells labeling Vimentin with Rabbit anti-Vimentin antibody (AWA10146) at 1/150 dilution(green). Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.03% Triton X-100 in PBS for 30 minutes, and then blocked with 5% BSA for 60 minutes at 37 ℃. Cells were then incubated with Rabbit anti-Vimentin antibody (AWA10146) at 1/150 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, AWS0005) was used as the secondary antibody at 1/200 dilution for 60 minutes at 37 ℃. Nuclear DNA was labelled in blue with DAPI(AWC0291).

	Fig：Immunoprecipitation of Vimentin from Hela cells was performed using Vimentin Rabbit mAb (AWA10146,1:250). Rabbit IgG isotype control was used to precipitate the Control IgG sample. The IP sample was eluted with Glycine buffer. Western blot analysis of immunoprecipitates was conducted using Vimentin Rabbit mAb (AWA10146) at a dilution of 1:1000. Goat Anti-Rabbit IgG(H+L) - HRP Secondary Antibody (AWS0002) at 1:5,000 dilution.

	Fig: Multiplex immunohistochemistry analysis of Rat-Myocardial muscle tissue (Formalin/PFA-fixed paraffin-embedded sections). Merged staining of anti-MYH1 (AWA10538; green; TSA-520:AWI0688), anti-Connexin 43(AWA10489; red; TSA-570: AWI0689), anti-Vimentin (AWA10146; white; TSA-690: AWI0691).Antibody Wash Solution (for mIHC) (AWI0707, 37℃, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.The section was incubated in three rounds of staining; in the order of MYH1 (AWA10538) (1/200 dilution), anti-Connexin 43 (AWA10489) (1/200 dilution), anti-Vimentin (AWA10146) (1/600 dilution); each using a separate fluorescent tyramide signal amplification system.DAPI (blue, AWC0291) was used as a nuclear counter stain.Microscopy and pseudocoloring of individual TSA dyes was performed using a pannoramic midi.

	Fig : Immunohistochemical analysis of paraffin-embedded Rat-heart tissue with Rabbit anti-Vimentin (AWA10146) at 1/800 dilution. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA10146) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system(ABIOWELL, AWI0629). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig : Immunohistochemical analysis of paraffin-embedded Mouse-heart tissue with Rabbit anti-Vimentin (AWA10146) at 1/800 dilution. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA10146) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system(ABIOWELL, AWI0629). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig：Immunoprecipitation of Vimentin from A549 cells was performed using Vimentin Rabbit mAb (AWA10146,1:250). Rabbit IgG isotype control was used to precipitate the Control IgG sample. The IP sample was eluted with Glycine buffer. Western blot analysis of immunoprecipitates was conducted using Vimentin Rabbit mAb (AWA10146) at a dilution of 1:1000. Goat Anti-Rabbit IgG(H+L) - HRP Secondary Antibody (AWS0002) at 1:5,000 dilution.

	Fig：Flow cytometric analysis of HELA cells labeling Vimentin. Overlay histogram showing HELA cells stained with Vimentin (green line). The cell were fixed in 4% paraformaldehyde for 30 minutes at 37 ℃, permeabilized with 0.02% Triton X-100 in PBS for 30 minutes,and then stained with the primary antibody(AWA10146, 1:50 ) for 30 min at 4°C. The secondary antibody used was an Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody (AWS0005b) at 1/2000 dilution for 30 min at 4ºC. Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

	Fig : Western blot analysis of Vimentin on different lysates. Proteins were transferred to a NC membrane and blocked with 5% NF-Milk in TBST for 1 hour at room temperature. The primary antibody (AWA10146, 1/1000) was used in TBST at room temperature for 2 hours. Goat Anti- Rabbit IgG - HRP Secondary Antibody (AWS0002) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: A549 cell Lane 2: PC-3 cell Lane 3: LN229 cell Lane 4: HELA cell Lane 5: Jurkat cell Lane 6: NRK-49F cell Lane 7: RBL-2H3 cell Predicted molecular weight:54 kDa Observed molecular weight:54 kDa

	Fig : Western blot analysis of Vimentin on different lysates. Proteins were transferred to a NC membrane and blocked with 5% NF-Milk in TBST for 1 hour at room temperature. The primary antibody (AWA10146, 1/1000) was used in TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (AWS0002) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control:* Lane 1: C2C12 cell Lane 2: HepG2 cell Predicted molecular weight:54 kDa Observed molecular weight:54 kDa

	Fig :Western blot analysis of Vimentin on different lysates. Proteins were transferred to a NC membrane and blocked with 5% NF-Milk in TBST for 1 hour at room temperature. The primary antibody (AWA10146, 1/1000) was used in TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (AWS0002) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control:* Lane 1: U251 cell Lane 2: Mouse heart Lane 3: C6 cell Predicted molecular weight: 54 kDa Observed molecular weight: 54 kDa