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GRK5 antibody - middle region (ARP54750_P050)

100 ul
$289.00
In Stock

Conjugation Options

ARP54750_P050-FITC Conjugated

ARP54750_P050-HRP Conjugated

ARP54750_P050-Biotin Conjugated

Gene Symbol:
GRK5
NCBI Gene Id:
2869
Official Gene Full Name:
G protein-coupled receptor kinase 5
Protein Name:
G protein-coupled receptor kinase 5
Swissprot Id:
P34947
Protein Accession #:
NP_005299
Nucleotide Accession #:
NM_005308
Alias Symbols:
GPRK5
Replacement Item:
This antibody may replace item sc-11396 from Santa Cruz Biotechnology.
Description of Target:
GRK5 is a member of the guanine nucleotide-binding protein (G protein)-coupled receptor kinase subfamily of the Ser/Thr protein kinase family. The protein phosphorylates the activated forms of G protein-coupled receptors thus initiating their deactivation. It has also been shown to play a role in regulating the motility of polymorphonuclear leukocytes (PMNs).This gene encodes a member of the guanine nucleotide-binding protein (G protein)-coupled receptor kinase subfamily of the Ser/Thr protein kinase family. The protein phosphorylates the activated forms of G protein-coupled receptors thus initiating their deactivation. It has also been shown to play a role in regulating the motility of polymorphonuclear leukocytes (PMNs). Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.
Protein Size (# AA):
590
Molecular Weight:
68kDa
Host:
Rabbit
Clonality:
Polyclonal
Purification:
Affinity Purified
Application:
WB, IHC
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express GRK5.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express GRK5.
Immunogen:
The immunogen is a synthetic peptide directed towards the middle region of human GRK5
Species Reactivity:
Cow, Dog, Guinea Pig, Horse, Human, Mouse, Rabbit, Rat, Yeast, Zebrafish
Predicted Homology Based on Immunogen Sequence:
Cow: 86%; Dog: 100%; Guinea Pig: 100%; Horse: 93%; Human: 100%; Mouse: 100%; Rabbit: 86%; Rat: 100%; Yeast: 79%; Zebrafish: 86%
Complete computational species homology data:
Anti-GRK5 (ARP54750_P050)
Peptide Sequence:
Synthetic peptide located within the following region: FYAAEILCGLEDLHRENTVYRDLKPENILLDDYGHIRISDLGLAVKIPEG
Product Format:
Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
Reconstitution and Storage:
For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
Concentration:
Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
Protein Interactions:
TBXA2R; USP47; C1QBP; YWHAB; XRCC5; UBTF; UBC; TUFM; NR2F1; TCOF1; SRPK1; SPTBN1; SPTAN1; SMARCA1; SKP1; SRSF1; RPL4; RPL3; RBMS1; PSMD11; PSMD2; PSMC6; PSMC2; PSMC1; PRKDC; PPP2R2A; PNN; PIP4K2A; PIP; NFIB; NCL; NCBP1; MYO9B; MYO1C; MYH10; MYH9; MYCN; MA
Blocking Peptide:
For anti-GRK5 (ARP54750_P050) antibody is Catalog # AAP54750 (Previous Catalog # AAPP31545)
Datasheets/Manuals:
Printable datasheet for anti-GRK5 (ARP54750_P050) antibody
Target Reference:
Liggett,S.B., (2008) Nat. Med. 14 (5), 510-517
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2 Item(s)

73/03/2018 16:36
  • Overall Experience:
  • Quality:
Mouse Heart in WB

Researcher: Kathleen Gabrielson

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02/01/2017 16:24
  • Overall Experience:
  • Quality:
Product Review: GRK5 antibody-middle region (ARP54750_P050) in Tobacco hornworm larvae intestine lysate using Western Blot
Product Page for GRK5 antibody-middle region (ARP54750_P050)

Researcher: Dra. Helena Porta Ducoing and Leidy Bedolla.
Departamento de Microbiología Molecular
Instituto de Biotecnología, UNAM

Application: Western blotting
Species + Tissue/Cell type: Tobacco hornworm larvae intestine lysate
How many ug's of tissue/cell lysate run on the gel:
1: 40 ug tobacco hornworm larvae intestine lysate
2: 40 ug tobacco hornworm larvae intestine lysate
3: 40 ug tobacco hornworm larvae intestine lysate
Primary antibody dilution: 1:1000
Secondary antibody: Goat anti-rabbit HRP
Secondary antibody dilution: 1:10000



Questionnaire:

How do Aviva's reagents play a role in your experimental goals?

It is very nice to use specific antibodies. Although the results is not clean we observed a band with the selected MW (40kD) for PKA kinase

How would you rate this antibody on a scale from 1-5 (5=best) and why?

4. The detection conditions most be modified.

Would you use this antibody in future experiments?

Yes

Have you used another antibody which has worked in your application?

No

Do you believe the information about the reagent on Aviva's website is correct?

Yes I think so

If the antibody works, do you plan to use it in future experiments or to publish your data?

Why or why not?

We are very interested I use this antibody

Sample Description:

Manduca sexta

Species, Tissue/Cell Type:

Larvae Intestine

How many different experimental trials were conducted using the antibody sample?

We made four trials using the same antibody dilution.

What type of experimental sample are you using and how did you preparing it?

We dissect intestine from larvae, homogenize the tissue in a cell lyses solution for WB

Primary used and dilution: 1:1000

Secondary used and dilution: 1:10000

Experimental Procedure/Protocols:

Aviva WB protocol

Immunoblotting

A. Reagents and Materials

•           Antibody dilution buffer:5% non-fat dry milk in 0.05% PBST

•           Developing solution/Fixative solution for X‐ray film

•           Membrane washing buffer: 0.1% PBST

•           Primary antibody; Aviva offers >30,000 antibodies for Western Blot

•           Secondary antibody; Goat anti‐Rabbit I gG-HRP (CAT#ASP00001)

•           Horseradish Peroxidase Substrate: e.g. Chemiluminescent FemtoMax (CAT# OORA01695)

•           X‐ray film

B. Equipment

•           Rotary shaker

C. Protocol

This protocol is to be followed after Gel Transfer.

•           Primary antibodies preparation

a) The final primary antibody concentration should be 1.25‐5.0ug/ml for protein A purified IgG and 0.25‐1.0ug/ml for peptide purified IgG.

•           Pretreat membrane: Wash the membrane in distilled water for 20min.

•           Primary antibodies incubation

•           a) Dilute the primary antibody in antibody dilution buffer to suitable antibody concentration, then incubation for 1hour at room temperature with agitation to enable adequate homogenous covering of the membrane and prevent uneven binding.

•           Wash membrane: Pour off primary antibody solution and wash membrane 6 times and for 5 minutes each time in Membrane washing buffer.

•           Secondary antibodies incubation

a) Dilute secondary antibody in antibody dilutionbuffer and incubate with shaking for 1 hour.

•           Wash membrane: Pour off primary antibody solution and wash membrane 6 times and for 5 minutes each time in Membrane washing buffer.

•           Development

a) Mix the two solutions of HRP substrate reagent in appropriate ratio immediately prior to adding to the membrane. Incubate with the membrane for 1 minute. Drain and remove excess reagent and place the membrane in plastic wrap to ensure a dry surface for film exposure.

•           Exposure in darkroom

a) Exposure the membranes to X‐ray film in cassette for 1 minute. Manual film development is used to control the incubation time of the x‐ray film in the developing solution and fixative solution. Normally it's about 2‐10 min for development and 5 min for fixation. Dry the film after thoroughly washing with water.

What controls were used in your experiment? Please include your positive control:

We do not have a constitutive control yet.

How did you store the antibody after re-suspension?

At -20 0C
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