IKBKG antibody - middle region (ARP30005_T100)

Catalog#: ARP30005_T100

• Size: 100ug

Human HepG2

Mouse testis and HeLa

• Description of Target: Familial incontinentia pigmenti (IP) is a genodermatosis that segregates as an X-linked dominant disorder and is usually lethal prenatally in males . In affected females it causes highly variable abnormalities of the skin, hair, nails, teeth, eyes, and central nervous system. The prominent skin signs occur in 4 classic cutaneous stages: perinatal inflammatory vesicles, verrucous patches, a distinctive pattern of hyperpigmentation, and dermal scarring. Cells expressing the mutated X chromosome are eliminated selectively around the time of birth, so females with IP exhibit extremely skewed X-inactivation. Familial incontinentia pigmenti is caused by mutations in the NEMO gene and is here referred to as IP2, or 'classical' incontinentia pigmenti. Sporadic incontinentia pigmenti, the so-called IP1, which maps to Xp11, is categorized as hypomelanosis of Ito
• Gene Symbol: IKBKG
• Official Gene Full Name: Inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase gamma
• Alias Symbols: IP; IP1; IP2; FIP3; IPD2; NEMO; FIP-3; Fip3p; AMCBX1; IKK-gamma
• Protein Accession#: NP_003630
• Nucleotide Accession#: NM_003639
• Swissprot Id: Q9Y6K9
• Host: Rabbit
• Clonality: Polyclonal
• Protein Size (# AA): 419
• Molecular Weight: 48kDa
• Application: WB
• Partner Proteins: BCL10, CYLD, IKBKB, IKBKG, PRKCQ, TAB3, TANK, TRAF3IP2, TRAF3IP2, UBB, ZFAND5, tax, BCL10, BIRC3, CARD10, CARD11, CARD8, CASP8, CDC37, CHUK, COPS3, CREBBP, CYLD, FADD, GSK3B, HSP90AA1, HSP90AB1, HSPA1A, HSPA4, IKBKB, IKBKG, MAP3K14, NCOA3, NFKB1, NFKB2, NFKBIA, NFKBIB, PPM1B, PRKCQ, RAB11B, RAL
• Immunogen: The immunogen for anti-IKBKG antibody: synthetic peptide directed towards the middle region of human IKBKG
• Product Format: Lyophilized powder
• Purification: Protein A purified
• Predicted Homology Based on Immunogen Sequence: Dog: 100%; Horse: 100%; Human: 100%; Rabbit: 100%; Guinea pig: 85%; Mouse: 85%; Pig: 85%; Rat: 85%
• Peptide Sequence: Synthetic peptide located within the following region: LGELQESQSRLEAATKECQALEGRARAASEQARQLESEREALQQQHSVQV
• Blocking Peptide: For anti-IKBKG antibody is Catalog # AAP30005 (Previous Catalog # AAPH00105)
• Key Reference: Hai,T., et al., (2006) J. Virol. 80 (9), 4227-4241
• Reconstitution and Storage: Add 100 μl of distilled water. Final anti-IKBKG antibody concentration is 1 mg/ml in PBS buffer. For longer periods of storage, store at -20°C. Avoid repeat freeze-thaw cycles.

Product Protocols: IKBKG antibody tested with Human Hepg2 Cells (ARP30005_T100)

Aviva Systems Biology is the original manufacturer of this IKBKG antibody (ARP30005_T100)

Click here to view the IKBKG antibody Western Blot Protocol

Product Datasheet Link: IKBKG antibody (ARP30005_T100)

WB Suggested Anti-IKBKG Antibody Titration: 1.25ug/ml
ELISA Titer: 1:312500
Positive Control: HepG2

Western Blot image:


Description of Target: Familial incontinentia pigmenti (IP) is a genodermatosis that segregates as an X-linked dominant disorder and is usually lethal prenatally in males . In affected females it causes highly variable abnormalities of the skin, hair, nails, teeth, eyes, and central nervous system. The prominent skin signs occur in 4 classic cutaneous stages: perinatal inflammatory vesicles, verrucous patches, a distinctive pattern of hyperpigmentation, and dermal scarring. Cells expressing the mutated X chromosome are eliminated selectively around the time of birth, so females with IP exhibit extremely skewed X-inactivation. Familial incontinentia pigmenti is caused by mutations in the NEMO gene and is here referred to as IP2, or 'classical' incontinentia pigmenti. Sporadic incontinentia pigmenti, the so-called IP1, which maps to Xp11, is categorized as hypomelanosis of Ito

Questions pertaining to this data can be directed to techsupport@avivasysbio.com

Aviva Systems Biology’s IKBKG antibody (ARP30005_T100) has been tested using other cell lysates and tissues. To obtain more data about this antibody please email us at info@avivasysbio.com.

To order by phone call us at (888) 880-0001, fax us at (858) 552-6975 or send an email to info@avivasysbio.com. Aviva manufactures this antibody so we can offer the best price. Please contact us to request pricing information.

All of Aviva’s products are guaranteed for the applications and experimental sample types mentioned in the datasheet below. Are you curious if this product will work for you? Please contact us at techsupport@avivasysbio.com

Product Review: IKBKG antibody - middle region (ARP30005_T100) in Mouse testis and HeLa cells using Western Blot

Product Page for IKBKG antibody - middle region (ARP30005_T100)

Researcher: Andreia Carvalho, Instituto de Biologia Molecular e Celular, Universidade do Porto (IBMC-UP)/Organelle Biogenesis and Function (OBF) Group
Application: Western blotting
Species+tissue/cell type: Mouse testis and HeLa cells
How many ug’s of tissue/cell lysate run on the gel:
1. 100 ug mouse testis lysate
2. 100 ug HeLa cell lysate
Primary antibody dilution: 1:800
Secondary antibody: Anti-rabbit-AP
Secondary antibody dilution: 1:10000

Questionnaire:

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

Our work has a strong component based on the isolation/purification of protein(s)/protein complexes from mammalian orign, relying mainly in biochemical approaches, and therefore the use of specific antibodies is of great importance to our studies.

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

2. The antibody seems to recognize a band corresponding to IKBKG (predicted molecular weight ~48kDa) in HeLa cells only.

Would you use this antibody in future experiments?

Only in HeLa cells.

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.

If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?

Yes. When working with protein biochemistry it is mandatory to have a specific antibody to validate the majority of our findings.

How did you store the antibody after re-suspension?

After resuspension, antibodies were divided into small aliquots and stored at -20ºC as described in your application sheet.

Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):

1)Mouse, human; 2) cytosolic factions from mouse organ and total homogenates from HeLa cells 3)100 micrograms of total protein in each lane.

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

3 with mouse cytosolic fractions from organs and HeLa cells.

How was this sample prepared?

Cytosolic fractions from mouse tissues were prepared as described in: Grou CP et al., Identification of ubiquitin-specific protease 9X(USP9X) as a deubiquitinase acting on ubiquitin-peroxin 5 (PEX5) thioester conjugate. JBC. 2012; 287:12815-27. Total homogenates from HeLa cells were prepared as described in: Pinto MP et al., Heat shock induces a massive but differential inactivation of SUMO-specific proteases. BBA-MCR. 2012; DOI: 10.1016/j.bbamcr.2012.07.010

SDS-PAGE:

100 micrograms of each sample were added to Laemmli sample buffer and incubated at 65ºC for 10 min followed by 5min at 95ºC. Following incubation, samples were centrifuged at Vmax for 2min and loaded into a 11% SDS-PAGE.

Primary antibody dilution and incubation time:

Dilutions tested: 1:800. Incubation time: overnight at 4ºC.

Secondary antibody used and dilution and incubation time:

Alkaline phosphatase-conjugated secondary antibody: anti-rabbit (SIGMA) dilution 1:10000. Incubation time: 1h at room temperature.

What controls were used in your experiment (positive/negative)?

None.

Please include your detailed WB Procedure/Protocol here:

All samples were run on an 11% SDS/PAGE and transferred to a nitrocellulose membrane.

Immunoblot.

Nitrocellulose membranes were stained with Ponceau S to confirm protein loadings. Following destaining with TBS1x, the membrane was blocked for 1h at room temperature (RT) with 5% skimmed milk(SM)/TBS1x. The blot was incubated overnight at 4ºC with a 1:1000 dilution of the mentioned antibody in 5% SM/TBS1x. Following a 2 wash step for 5min at RT with 1X TBS-T(tween 20 0.1%), the membrane was incubated with a alkaline phosphatase-conjugated secondary antibody (anti-rabbit, Sigma; dilution 1:10000) for 1h at RT in 5% SM/TBS1x. Again a 2 wash step for 5min at RT with 1X TBS-T(tween 20 0.1%) was performed and the membrane was incubated with the developing agents NBT/BCIP (BioRad).