Antibodies, Proteins, Kits and Reagents for Life Science

371 894 ₸ Product size

100 µl 371 894 ₸

Order now and get it on Tuesday March 02, 2021

Anti-ROCK1 antibody [EPR638Y] (ab134181)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR638Y] to ROCK1
Suitable for: WB, IP, IHC-P
Knockout validated
Reacts with: Human

Product name

Anti-ROCK1 antibody [EPR638Y]
See all ROCK1 primary antibodies
Description

Rabbit monoclonal [EPR638Y] to ROCK1
Host species

Rabbit

Tested Applications & Species

Application	Species
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human ROCK1 aa 1300-1400 (C terminal). The exact sequence is proprietary.
Positive control
- WB: HeLa cell lysate, HEK293 cell lysate; IHC: Human lymphoma tissue; IP: HeLa cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR638Y
Isotype

IgG
Research areas

Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)

Lane 1: Wild type HAP1 whole cell lysate (40 µg)
Lane 2: ROCK1 knockout HAP1 whole cell lysate (40 µg)
Lane 3: HEK293 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab134181 observed at 165 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab134181 was shown to specifically react with ROCK1 when ROCK1 knockout samples were used. Wild-type and ROCK1 knockout samples were subjected to SDS-PAGE. Ab134181 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EPR638Y] (ab134181)

Immunohistochemical analysis of paraffin embedded Human lymphoma tissue labelling ROCK1 with ab134181 at a dilution of 1/50.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)

All lanes : Anti-ROCK1 antibody [EPR638Y] (ab134181) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ROCK1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 158 kDa
Observed band size: 160 kDa
why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab134181 observed at 160 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab134181 was shown to react with ROCK1 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264780 (knockout cell lysate ab257642) lane below 160kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and ROCK1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab134181 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunoprecipitation - Anti-ROCK1 antibody [EPR638Y] (ab134181)

Purified ab134181 at 1/120 dilution (2µg) immunoprecipitating ROCK1 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab134181 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab134181 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 158 kDa
Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)

Anti-ROCK1 antibody [EPR638Y] (ab134181) at 1/500 dilution + HeLa cell lysate at 10 µg

Secondary
HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 158 kDa
Anti-ROCK1 antibody [EPR638Y] (ab134181)

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